DETAILED OFFICE ACTION
Status of the Application
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Applicant’s amendment to the claims, filed on September 18, 2025, is acknowledged. This listing of the claims replaces all prior versions and listings of the
claims.
Claims 16-37 are pending and are being examined on the merits.
Claims 1-15 are cancelled.
Applicant’s amendment to the specification, filed September 18, 2025, is acknowledged.
Applicant’s remarks filed September 18, 2025 in response to the non-final rejection mailed June 27, 2025, is acknowledged and have been fully considered.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Election/Restrictions
In response to a requirement for restriction/election mailed March 13, 2025,
applicant elected with traverse Group 1 (claims 16-18, 20-21, and 23-29) and the following species in the reply filed May 8, 2025:
Species Group 1) Applicant elects as impact of a fusion protein on a protein of interest: Accumulation;
Species Group 2) Applicant elects as the peptide of interest: matricin proteins; and
Species Group 3) Applicant elects as the alga: Chlamydomonas.
The requirement of restriction was proper. However, examiner withdrew the restriction requirement to elect a single group of claims but maintained the requirements for species elections. The requirement for a species election for Species is also now withdrawn by the examiner. Therefore, claims 16-37 and elected species are now under examination.
Specification Objections
In view of the amendments to the specification filed September 18, 2025, the objection to specification is withdrawn with regard to the following informalities: The reference to sequence identifiers as “SEQ ID N°:” or “SEQ ID Nos:”.
Claim Objections
Claim 18 is objected to in the recitation of “and optionally; and” and in the interest of improving claim form, it is suggested that the noted phrase be amended to delete the second instance of “and.”
Claim 20 is objected to in the recitation of “in comparison with the production in algae without aprotinin” and in the interest of improving claim form, it is suggested that the noted phrase be amended to recite “in comparison with the recombinant peptide, polypeptide or protein of interest produced in algae that is not fused to aprotinin.”
Claim 31 is objected to in the recitation of “matricins proteins, polypeptides or peptides” and in the interest of improving claim form and consistency with claim 22, it is suggested that the noted phrase be amended to delete “proteins, polypeptides or peptides.”
Claim Rejections - 35 USC § 112(b)
The rejection of claims 16-18, 20-21, and 23-29 under 35 U.S.C. 112(b) for being indefinite because the recitation of the phrase “transforming transformation” is unclear is withdrawn in view of the amendment to claim 16 to remove “transformation of.”
The rejection of claim 18 under 35 U.S.C. 112(b) for being indefinite because the claim recites the term "microalgae" which lacks antecedence is withdrawn in view of the amendment to modify “microalgae” to “algae.”
The rejection of claim 20 under 35 U.S.C. 112(b) for being indefinite because the term "microalgae" in lines 3-4 lacks antecedence is withdrawn in view of the amendment to the claim to remove the term “microalgae.”
The rejection of claim 20 under 35 U.S.C. 112(b) for being indefinite because the term "accumulation" in lines 3-4 lacks antecedence is withdrawn in view of amending claim 20 to an independent claim.
The rejection of claim 20 under 35 U.S.C. 112(b) is rejected for being as indefinite because of the use of exemplary claim language in the phrase “in algae, in particular in microalgae, more particularly in the chloroplast of microalgae” is withdrawn in view of the amendment of claim 20 to remove the exemplary language.
The rejection of claim 20 under 35 U.S.C. 112(b) for being indefinite because the use of “proteins peptide” is unclear as to the intended meaning in the phrase “of recombinant proteins peptide, polypeptide or protein of interest in algae” is withdrawn in view of the amendment to the claim to remove “proteins” from “proteins peptide.”
The rejection of claim 20 under 35 U.S.C. 112(b) for being indefinite because of the use of relative term “increase” in the phrase “to increase accumulation” is withdrawn in view of the amendment to the claim to provide a reference point to determine increased accumulation.
The rejection of claims under 35 U.S.C. 112(b) 22-23 for being indefinite because the use of the plural term “matricins” in the phrase “matricins proteins, polypeptides or peptides” is unclear is withdrawn in view of the amendment to the claims to modify the phrase “matricins proteins, polypeptides or peptides” to “matricins.”
The rejection of claim 21-23 and 31 under 35 U.S.C. 112(b) for being indefinite for reciting a broad and narrow narrower statement of the limitation is withdrawn upon further consideration of the rejection.
Claims 21 and 30-31 are rejected under 35 U.S.C. 112(b) as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor regards as the invention.
The rejection of claims 21 and 31 under 35 U.S.C. 112(b) for being indefinite because the use of the plural term “matricins” in the phrase “matricins proteins, polypeptides or peptides” is unclear is maintained. While there are a plurality of instances in the specification that use the term “matricins”, ending the term with an “s” suggests a plurality of the moieties and a plurality of them would not be considered by an ordinary artisan to be a recombinant protein, polypeptide or peptide of interest as described in claims 16 and 19 as well as claims dependent therefrom. The specification uses a singular version of “matricin” on p. 10, para 2 when it recites, “elastin peptides are part of matricin peptides” suggesting the plural form of the word is a typographical error. It is also not clear whether the phrase “proteins, polypeptides or peptides” refer to the matricin itself or whether these “proteins, polypeptides or peptides” are distinct from matricin. For the purpose of compact prosecution, the limitation of “matricins” in the claims is construed to refer to “matricin” and the “proteins, polypeptides or peptides” are considered to belong to matricin. Claim 30 is newly rejected as indefinite for reciting a “use” without setting forth any method step(s) for carrying out the method. It is unclear as to how the “recombinant algae according to claim 19” is to be used to produce a fusion protein. The Office recommends amending the claim to recite a method step or steps for producing a fusion protein with the “recombinant algae according to claim 19,” e.g., “said method comprising culturing the recombinant algae according to claim 19 to produce the fusion protein.”
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
Claims 19-20, 30 and 35 are newly rejected under 35 U.S.C. 102(a)(1) as being anticipated by Paget et al. (WO 2008/003666 A1; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Paget”).
The claims are drawn to recombinant algae comprising a nucleic acid sequence encoding a fusion protein, wherein said fusion protein comprises aprotinin and a recombinant protein, polypeptide or peptide of interest as well as the use thereof to produce a fusion protein.
Regarding claims 19-20, 30 and 35, Paget teaches a method of expressing a recombinant chimeric protein of interest in the thylakoid lumen of plants comprising transforming the cells with a vector encoding a bacterial signal peptide fused with a heterologous nucleic acid sequence encoding a recombinant or protein or peptide of interest (Abstract; p. 3, ln 22-25, p. 4, ln 1-8; p. 11, ln 5-16, p. 11, ln19-22; p. 13, ln 31 – 33; p. 14, ln 8-15; p. 17, ln 31 – p. 18, ln 18; Claims 1-2, 5, 7-9, and 13-14). Paget teaches the transplastomic plant and/or progeny can comprise algae (p. 13, ln 31-33). The method leads to high-level accumulation of active and correctly folded recombinant enzyme in the thylakoid lumen of chloroplasts (Abstract; p. 23, ln 26- p. 24, ln 15).
For the reasons stated herein, claims 19-20, 30 and 35 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Paget.
Claim Rejections - 35 USC § 103
The rejection of claims 16-20, 28, and 35 under 35 U.S.C. 103 as being unpatentable over Paget in view of Collier et al. (US 2005/0202535A1; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Collier”) and Itzhaki et al. (Enzymology, published March 1998, Vol. 273, No. 12, p. 7094-7098; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Itzhaki”) is maintained.
The claims are drawn to a method for producing a recombinant protein, polypeptide, or peptide of interest in algae, wherein the method comprises transforming algae with a specific nucleic acid sequence encoding a fusion protein, wherein said fusion protein comprises a protein and said recombinant protein, polypeptide or peptide of interest.
Regarding claims 16-17, 19-20, 28, and 35,
Paget teaches a method of expressing a recombinant chimeric protein of interest in the thylakoid lumen of plants comprising transforming the cells with a vector encoding a bacterial signal peptide fused with a heterologous nucleic acid sequence encoding a recombinant or protein or peptide of interest (Abstract; p. 3, ln 22-25, p. 4, ln 1-8; p. 11, ln 19-22; p. 13, ln 31 – 33; p. 14, ln 8-15; p. 17, ln 31 – p. 18, ln 18; Claims 1-2, 5, 7-9, and 13-14). Paget teaches the fusion protein comprising a bacterial peptide is able to translocate across the thylakoid membrane and localize to the lumen of thylakoids in plants (p. 22, ln 26 – p. 24, ln 15). Paget teaches the transplastomic plant and/or progeny can comprise algae (p. 13, ln 31-33). The method leads to high-level accumulation of active and correctly folded recombinant enzyme in the thylakoid lumen of chloroplasts (Abstract; p. 23, ln 26- p. 24, ln 15). The thylakoid lumen of chloroplasts is a cellular compartment optimal for the accumulation of certain recombinant proteins due to its content of proteases (p. 2, ln 25-27). Paget teaches the peptide of interest can be aprotinin and teaches aprotinin can be fused to a protein of interest as well as the petA (i.e. petA:aprotinin) (p. 2, ln 31 – p. 3, ln 6; p. 11, ln 5-16; p. 15, ln 35 - p. 16, ln 2). Aprotinin is known to inhibit serine proteases including trypsin, chymotrypsin, plasmin and kallikrein (p. 11, ln 6-9). Paget teaches constructing a transformation vector by cloning a nucleic acid sequence of interest into an expression vector (p. 17, ln 31 – p. 18, ln 19).
Paget does not teach wherein the chimeric protein that comprises a signal peptide and aprotinin also comprises an additional protein, polypeptide, or peptide of interest.
Collier teaches a fusion protein comprising a protease inhibitor and a peptide of interest (Claim 1). In one embodiment, the fusion protein consists in order of a signal sequence, a peptide of interest, a cleavable linker region, and a protease inhibitor or variant thereof (para [0016]).
Itzhaki teaches proteolytic degradation of proteins in the thylakoid lumen, but discloses that the proteases involved are largely unknown (Abstract; p. 7094, col 2, para 2). Apolastocyanin is an example of a protein transported across the thylakoid membrane in the thylakoid lumen of Chlamydomonas cells but it fails to accumulate due to rapid degradation (p. 7094, col 2, para 2). Serine protease DegP is implicated in the proteolytic degradation of proteins in the thylakoid lumen (Abstract). Isolated thylakoid membranes containing the chloroplast homologue of DegP degraded b-casein, an in vitro substrate of the bacterial protease; this activity was partially inhibited by a serine protease inhibitor, suggesting that at least part of the casein-degrading activity in the thylakoid membrane is attributable to the serine proteases, such as DegP (Abstract). Based on these teachings, an ordinary artisan would immediately recognize that the thylakoid lumen of chloroplasts contains at least one serine protease that degrade proteins as well as the activity of the serine protease can be inhibited by serine protease inhibitors.
In view of the combined teachings of Paget, Collier, and Itzhaki, it would have been obvious for a person of ordinary skill in the art before the effective filing date of the claimed invention to modify the method of expressing a recombinant chimeric protein of interest in the thylakoid lumen algae chloroplasts comprising transforming the cells with a vector encoding a bacterial signal peptide fused with a heterologous nucleic acid sequence encoding a recombinant or protein or peptide of interest taught by Paget such that the protein or peptide of interest comprises the fusion protein comprising a protease inhibitor and a peptide of interest taught by Collier and that the protease inhibitor comprises aprotinin taught by Paget, thereby arriving at the invention of claims 16-17, 19-20, 28, and 35.
An ordinary artisan would have been motivated to modify the nucleic acid encoding a signal peptide fused with a protein or peptide of interest in the method of producing a peptide of interest in the thylakoid lumen of algae taught by Paget such that the peptide of interest comprises the fusion protein comprising a protease inhibitor and a peptide of interest taught by Collier further substituted wherein the protease inhibitor comprises aprotinin – for the advantage of decreasing proteolytic degradation of the chimeric protein and/or the protein of interest in the thylakoid lumen of chloroplasts and therefore increasing the accumulation of the chimeric protein and/or the protein of interest. This is because Itzhaki taught the proteolytic degradation of proteins in the thylakoid lumen and implicated the role of serine proteinase DegP which can be partially inhibited by a serine protease inhibitor, and Paget taught aprotinin is a serine protease inhibitor. Therefore, the fusion of aprotinin to the protein of interest would provide the advantage of inhibiting serine proteases in the thylakoid lumen of chloroplasts and result in the increased accumulation of the chimeric protein and/or the protein of interest in the thylakoid lumen of chloroplasts.
An ordinary artisan would have had a reasonable expectation of success modifying the nucleic acid encoding the protein or peptide of interest taught by Paget to comprise the fusion protein comprising a protease inhibitor and a peptide of interest taught by Collier further substituted such that the protease inhibitor comprises aprotinin. This is because Paget already taught the chimeric protein comprising aprotinin and that aprotinin is a serine proteinase inhibitor, and Collier already taught a fusion protein comprising a protease inhibitor and a peptide of interest. A simple substitution of one polypeptide of interest for another polypeptide of interest, such as a fusion protein of interest, would have been expected to yield predictable results. An ordinary artisan would have had a reasonable expectation of success yielding increased accumulation of recombinant proteins peptide, polypeptide or protein of interest in algae because the modified polypeptide of interest comprising aprotinin would have inhibited serine proteases in the thylakoid lumen of chloroplast. This is because Itzhaki taught the proteolytic degradation of proteins in the thylakoid lumen and implicated the role of serine proteinase DegP which can be partially inhibited by a serine protease inhibitor, and Paget taught the fusion protein can comprise aprotinin which is a serine protease inhibitor. Therefore, the fusion of aprotinin to the protein of interest would provide the advantage of inhibiting serine proteases in the thylakoid lumen of chloroplasts and result in the increased accumulation of the chimeric protein and/or the protein of interest in the thylakoid lumen of chloroplasts.
Regarding claim 18, the modified method of producing a recombinant protein comprising transforming algae cells with a vector comprising a chimeric fusion protein comprising aprotinin and a peptide of interest taught by the combination of Paget, Collier, and Itzhaki, as applied to claims 16-17, 19-20, 28, and 35, are discussed above and incorporated herein.
Regarding claim 18(iv), Paget further teaches a selective marker that confers to a cell resistance against a chemical agent in order to select for transformed cells which have incorporated a chimeric gene (p. 12, ln 24-25).
Regarding claim 18(v), Paget teaches the analysis of proteins of transformed and untransformed cells via western blot analysis and two-dimensional gel electrophoresis (p. 20, ln 8 – p. 21, ln 9). Paget also teaches characterizing host cells expressing a protein of interest via in vitro and in vivo assays (p. 22, ln 12-30; meeting the claim limitations of dependent claim 18(v)).
Regarding claim (vi), Paget teaches extraction of a protein of interest from a transplastomic plant cell (p. 16, ln 25 – p. 17, ln 7; p. 20, ln 8 – p. 21, ln 9; Claim 13-14). Paget teaches transplastomic plant and/or progeny can comprise algae (p. 13, ln 31- p. 2).
In view of the combined teachings of Paget, Collier, and Itzhaki, it would have been obvious for a person of ordinary skill in the art before the effective filing date of the claimed invention to modify the method of producing aprotinin fusion protein in algae taught by the combination of Paget, Collier, and Itzhaki wherein the method further comprises identifying the transformed algae host cell, characterizing the microalgae host cell for the production of recombinant fusion protein, and extracting the recombinant fusion protein, thereby arriving at the invention of claim 18.
Since Paget teaches selective markers use to select for transformed cells that have incorporated a chimeric gene, an ordinary artisan would have been motivated to include a selectable marker in the vector used to transform algae in the method of producing recombinant protein taught by the combination of Paget, Collier, and Itzhaki in order to identify, cultivate, or process only algae that are transformed and thus capable of producing the aprotinin fusion protein to maximize the amount of fusion protein produce. Since Paget teaches analyzing transformed host cells for the production of a protein of interest via Western blot analysis and two-dimensional electrophoresis, an ordinary artisan would have been motivated to characterize the aprotinin fusion protein in the algae host cell taught by the combination of Paget, Collier, and Itzhaki in order to identify if the protein of interest is being produced. Since Paget teaches extracting a produced peptide of interest of interest from plants and algae cells, an ordinary artisan would have been motivated to extract the aprotinin fusion protein of interest for downstream applications of interest such as evaluation via Western blot analysis or two-dimensional gel electrophoresis.
An ordinary artisan would have had a reasonable expectation of success of introducing a selective marker into vector comprising the aprotinin fusion protein and selecting for cells that are transformed since Paget taught the use selective markers to select for transformed cells that have incorporated a chimeric gene. An ordinary artisan would have had a reasonable expectation of success of characterizing the microalgae host cell for the production of recombinant fusion protein since Paget teaches analyzing transformed host cells for the production of a protein of interest via Western blot analysis and two-dimensional electrophoresis. An ordinary artisan would have had a reasonable expectation of success of extracting the aprotinin fusion protein of interest because Paget teaches extracting a produced peptide of interest of interest from plants and algae cells.
Consequently, the invention of claims 16-20, 28, and 35 would have been obvious to one of ordinary skill in the art before the effective filing date.
The rejection of claims 24-25, 32, and 36-37 under 35 U.S.C. 103 as being unpatentable over Paget in view of Collier and Itzhaki, as applied to claims 16-20, 28, and 35 above, and further in view of Baudelet et al. (Algal Research, published July 2017, Vol. 25, p. 333-371; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Baudelet”) is maintained. Newly added claims 36-37 are included in the rejection and addressed by evidentiary reference Coll (Spanish Journal of Agricultural Research, published 2006, Vol. 4, No. 4, p. 316-330; cited on the attached Form PTO-892; hereafter “Coll”), which is necessitated by the addition of claims 36-37 and the new limitation requiring “microalgae” in the method.
Regarding claims 24-25, 32, and 36-37, the relevant teachings of Paget, Collier, and Itzhaki, as applied to claims 16-20, 28, and 35, are discussed above and incorporated herein.
Paget further teaches that Chlamydomonas reinhardtii is a single celled alga (p. 12, ln 12).
Paget does not explicitly teach wherein the algae in the method is Chlamydomonas nor that Chlamydomonas is a subset of algae classified as Chlorophyta.
Baudelet teaches that the algae Chlamydomonas reinhardtii is a type of Chlorophyta (p. 334, col 2, para 1; p. 335, col 1, para 2).
Coll is cited in accordance with MPEP 2131.01.III to show Chlamydomonas reinhardtii is a microalgae (Table 1; p. 316, col. 1, para 1). Therefore, the algae Chlamydomonas reinhardtii taught by Baudelet is inherently microalgae.
In view of the combined teachings of Paget, Collier and Itzhaki, and Baudelet, it would have been obvious for a person of ordinary skill in the art before the effective filing date of the claimed invention that the method of producing aprotinin-based fusion protein in algae taught by the combination of Paget, Collier and Itzhaki utilize the algae from Chlamydomonas taught by both Paget and which Baudelet teaches is a type of Chlorophyta, thereby arriving at the invention of claim 24-25 and 32. An ordinary artisan would have been motivated to use algae that are classified as a Chlamydomonas to produce aprotinin-based fusion proteins since Paget teaches the alga Chlamydomonas reinhardtii, and a simple substitution of one known type of alga for another type of alga would have been expected to generate similar and predictable results. Since Baudelet teaches the alga Chlamydomonas is a type of Chlorophyta, an ordinary artisan would have been motivated to use an alga classified as Chlorophyta for the same reason.
An ordinary artisan would have had a reasonable expectation of using an alga classified as Chlamydomonas to express the aprotinin-based fusion protein taught by the combination of Paget, Collier and Itzhaki because Paget teaches the alga Chlamydomonas reinhardtii, and a simple substitution of one known type of alga for another type of alga would have been expected to generate similar and predictable results. Since Baudelet teaches the alga Chlamydomonas is a type of Chlorophyta, an ordinary artisan would have had a reasonable expectation of using algae classified as Chlorophyta to express aprotinin-based fusion protein for the same reason.
Consequently, the invention of claims 24-25, 32, and 36-37 would have been obvious to one of ordinary skill in the art before the effective filing date.
The rejection of claims 26 and 33 under 35 U.S.C. 103 as being unpatentable over Paget in view of Collier and Itzhaki, as applied to claims 16-20, 28, and 35 above, and further in view of Chen et al. (Advanced Drug Delivery Reviews, published October 15, 2013, Vol. 65, NO 10, p. 1357-1369; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Chen”) is maintained.
Regarding claims 26 and 33, the relevant teachings of Paget, Collier, and Itzhaki, as applied to claims 16-20, 28, and 35, are discussed above and incorporated hereafter.
The combined teachings of Paget, Collier, and Itzhaki do not teach or suggest that in an aprotinin-based fusion the protein of interest is fused to the C-terminus of aprotinin.
Chen teaches creating fusion proteins comprising in all possible directions (i.e., (Protein A)-(Protein B) and (Protein B)-(Protein A)) in order to test the optimal orientation (p. 1363, col 2, para 3).
In view of the combined teachings of Paget, Collier, Itzhaki, and Chen, it would have been obvious for a person of ordinary skill in the art before the effective filing date of the claimed invention to modify the fusion protein comprising aprotinin and a protein or peptide of interest taught by the combination of Paget, Collier, Itzhaki such that the protein or peptide of interest is fused to the C-terminus of aprotinin, thereby arriving at the invention of claim 26 and 33. An ordinary artisan would have been motivated to fuse the protein or peptide of interest the C-terminus of aprotinin in order to determine the most effective orientation of the fusion protein since Chen taught producing fusion proteins in a plurality of orders in to determine the optimal orientation of the peptides within the fusion protein.
An ordinary artisan would have had a reasonable expectation of success of fusing a protein or peptide of interest to the C-terminus of aprotinin since Chen taught producing fusion proteins in a plurality of orders in to determine the optimal orientation of the peptides within the fusion protein.
Consequently, the invention of claims 26 and 33 would have been obvious to one of ordinary skill in the art before the effective filing date.
The rejection of claim 29 under 35 U.S.C. 103 as being unpatentable over Paget in view of Collier and Itzhaki, as applied to claims 16-20, 28, and 35 above, and further in view of Chen et al. (Advanced Drug Delivery Reviews, published October 15, 2013, Vol. 65, NO 10, p. 1357-1369; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Chen”) is maintained.
Regarding claim 29, the teachings of Paget, Collier, and Itzhaki, as applied to claims 16-20, 28, and 35, are discussed above and incorporated hereafter.
Collier further teaches a fusion protein comprising a protease inhibitor and a peptide of interest (Claim 1). In one embodiment, the fusion protein consists in order of a signal sequence, a peptide of interest, a cleavable linker region, and a protease inhibitor or variant thereof (para [0016]).
The combined teachings of Paget, Collier, and Itzhaki do not explicitly teach or make obvious wherein the fusion protein also comprises a protease specific cleavage site between the aprotinin and the recombinant protein or peptide of interest.
Chen teaches protease specific cleavable linkers between domains of fusion proteins (Abstract; p. 1361, col 2, para 3; Tables 2-3, Figure 2-4). Cleavable linkers enable targeting but also release a free functional domain of a fusion protein after cleavage (Abstract; Table 3).
In view of the combined teachings of Paget, Collier, Itzhaki, and Chen, it would have been obvious for a person of ordinary skill in the art before the effective filing date of the claimed invention that the aprotinin fusion protein comprising aprotinin and a protein or peptide of interest made Paget, Collier, Itzhaki, and Chen to comprise between the aprotinin and the protein, polypeptide, or peptide of interest a protease specific cleavable linker taught by Chen, thereby arriving at the invention of claim 29.
An ordinary artisan would have been motivated to modify the aprotinin fusion protein comprising aprotinin and a protein or peptide of interest made Paget, Collier, Itzhaki such that it comprises between the aprotinin and the protein or peptide of interest a protease specific cleavable linker in order to free the protein or peptide of interest from the fusion protein for isolation. This is because Collier taught a fusion protein comprising a cleavable linker between a protein or peptide of interest and a protease inhibitor and Chen taught protease specific cleavable linkers as well as that cleavable linkers allow for the release a free functional domain of a fusion protein after cleavage.
An ordinary artisan would have had a reasonable expectation of success of modifying the aprotinin fusion protein comprising aprotinin and a protein or peptide of interest made Paget, Collier, Itzhaki to comprise between the aprotinin and the protein or peptide of interest a protease specific cleavable linker because Chen taught the use of protease specific cleavable linkers between domains of fusion proteins to free a functional domain of the fusion protein after cleavage.
Consequently, the invention of claim 29 would have been obvious to one of ordinary skill in the art before the effective filing date.
The rejection of claims 27 and 34 under 35 U.S.C. 103 as being unpatentable over Paget in view of Collier and Itzhaki, as applied to claims 16-20, 28, and 35 above, and further in view of Mayfield et al. (US 2017/0015745 A1; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Mayfield”) is maintained.
Regarding claims 27 and 34, the relevant teachings of Paget, Collier, and Itzhaki, as applied to claims 16-20, 28, and 35, are discussed above and incorporated hereafter.
The combined teachings of Paget, Collier, and Itzhaki do not teach or suggest wherein the fusion protein comprises an epitope tag.
Mayfield teaches the expression of fusion protein in algae comprising a FLAG epitope tag for the purpose of analyzing the expression of the protein as well as for using affinity purification (Title; Abstract; para [0155, 0174]; Claim 21). The algae can comprise Chlamydomonas reinhardtii (para [0032]).
In view of the combined teachings of Paget, Collier, Itzhaki, and Mayfield, it would have been obvious for a person of ordinary skill in the art before the effective filing date of the claimed invention that the fusion protein taught by the combination of Paget, Collier, and Itzhaki comprise an epitope tag such as the FLAG epitope tag taught by Mayfield, thereby arriving at the invention of claims 27 and 34. An ordinary artisan would have been motivated to incorporate an epitope tag into the fusion protein taught by the combination of Paget, Collier, and Itzhaki in order to analyze or purify the fusion protein because Mayfield taught the use of a FLAG epitope tag in a fusion protein produced in algae for the purpose of analyzing the expression of the protein as well as for using affinity purification.
An ordinary artisan would have had a reasonable expectation of success of incorporating an epitope flag into the fusion protein taught by the combination of Paget, Collier, and Itzhaki because Mayfield taught the use of a FLAG epitope tag in a fusion protein produced in algae for the purpose of analyzing the expression of the protein as well as for using affinity purification.
Consequently, the invention of claims 27 and 34 would have been obvious to one of ordinary skill in the art before the effective filing date.
The rejection of claims 21-23 and 31 under 35 U.S.C. 103 as being unpatentable over Paget in view of Collier and Itzhaki as applied to 16-20, 28, and 35 above, and further in view of Stutz et al. (US 2010/0316720 A1; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Stutz”), Gunin et al. (Advances in Gerontology, published January 5, 2012, Vol. 1, p. 299-303; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Gunin”), and Koria et al. (PNAS, published January 18, 2011, Vol. 18, No. 3, p. 1034-1039; cited on the Form PTO-892 mailed 6/27/2025; hereafter “Koria) is maintained.
Regarding claims 21-23 and 31, since claims 23 and 31 define elastin and elastin like proteins as a type of matricin proteins, elastin like proteins are interpreted as matricin in claims 21-23.
The teachings of Paget, Collier, and Itzhaki, as applied to claims 16-20, 28, and 35, are discussed above and incorporated here.
The combined teachings of Paget, Collier, and Itzhaki do not teach or suggest that the protein of interest is chosen from matricin proteins, polypeptides or peptide nor wherein the said matricin proteins, polypeptides or peptides are chosen from elastin and elastin like proteins.
Stutz teaches the production of algae extracts and its use in cosmetics and pharmaceuticals to guard against extrinsic aging of the skin (Abstract). Application of extract inhibits the expression of matrix metalloproteinases 1 and 3, and matrix metalloproteinases 1 and 3 are higher in aged skin and responsible for the depletion of structure proteins like elastin and which play a role in skin aging (para [0026]).
Gunin teaches the total number of fibroblasts in skin decreases progressively with age and that the reduced number of fibroblasts is partly due to the reduced activity of proliferation in the cells (Abstract).
Koria teaches elastin-like polypeptides (termed ELPs) increase fibroblast proliferation (p. 1034, col 2, para 2). Koria teaches a fusion protein comprising ELPs (Abstract).
In view of the combined teachings of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria it would have been obvious for a person of ordinary skill in the art before the effective filing date of the claimed invention to modify the fusion protein comprising aprotinin and a protein or peptide of interest taught by the combination of Paget, Collier, Itzhaki such that the polypeptide or peptide of interest comprise an ELP taught by Koria, thereby arriving at the inventions of claim 21 and 23. An ordinary artisan would have been motivated to modify the fusion protein taught by the combination of Paget, Collier, Itzhaki comprising aprotinin and a protein or peptide of interest such the protein or peptide of interest comprises ELP so that the extract of the algae comprising the aprotinin-ELP fusion can be used in cosmetics or pharmaceutics to treat aging because Stutz taught the production of algae extracts and its use in cosmetics and pharmaceuticals to guard against aging of skin, Gunin taught the total number of fibroblasts in skin decreases progressively with age and that the reduced number of fibroblasts is partly due to the reduced activity of proliferation in the cells, and Koria teaches ELPs that increase the proliferation of fibroblasts.
An ordinary artisan would have had a reasonable expectation of success of modifying the fusion protein comprising aprotinin and a protein, polypeptide, or peptide of interest taught by the combination of Paget, Collier, Itzhaki such that the polypeptide or peptide of interest comprise an ELP because Koria taught a fusion protein comprising ELP.
Consequently, the invention of claim 21-23 and 31 would have been obvious to one of ordinary skill in the art before the effective filing date.
RESPONSE TO REMARKS: Applicant submits that the instant claims are not disclosed or suggested by the cited prior art, taken either singly or in combination as well as that the instant claims are the result of a surprising and unexpected beneficial result. Particularly, that there is no disclosure or suggestion in the Paget, Collier, and Itzhaki references, taken either singly or in combination, to fuse aprotinin with a recombinant protein to express in algae and no motivation for the skilled artisan to combine these teachings with the goal to improve accumulation of recombinant proteins in an algae system as in the present claims. Further, the cited environments (bacterial vs. algal plastids) differ substantially so that one skilled in the art would have no reasonable expectation of success. Still further, as shown in Applicant's specification and as pointed out above, the use of aprotinin in the context of the present claims provided a surprising and unexpected beneficial result:, namely the accumulation of recombinant proteins in the algae system.
Applicant’s arguments are not found persuasive. It would have been obvious to one of ordinary skill in the art to modify the claimed method of expressing a recombinant chimeric protein of interest in the thylakoid lumen of algae comprising aprotinin from Paget with the recited prior art such that that the aprotinin comprises a fusion protein with a protein of interest in order to increase the accumulation of the protein of interest and/or the fusion protein comprising thereof as well as meets all of the claim limitations of the instant claims; therefore, there is no surprising and unexpected beneficial result. Furthermore, Paget teaches a fusion protein comprising a bacterial peptide is able to translocate across the thylakoid membrane and localize to the lumen of thylakoids in plants (p. 22, ln 26 – p. 24, ln 15). Since Paget teaches transplastomic plant and/or progeny can comprise algae, there is reasonable expectation of success in expressing the fusion protein taught by the recited art in the thylakoid lumen of algae.
Double Patenting Rejections
The previous provisional rejection of claims 16-35 for being provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-4, 6, 10-14 of copending Application No. 17/802,560 in view of Paget, Collier, Itzhaki, Chen, Mayfield, Stutz, Gunin, and Koria is withdrawn and a new rejection provided below that also includes newly added claims 36-37.
Claims 16-25, 28, 30-32, and 35 are newly provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim 2-4, 11, and 13-14 of copending Application No. 17/802,560 (hereafter, “App’560”) in view of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria.
Regarding claims 16, 19-23, 30-31, and 36-37, Claim 2 of App’560 is drawn to a method for producing a recombinant protein, polypeptide or peptide comprising repeat units of amino acids in the chloroplast of microalgae, said protein, polypeptide or peptide being chosen from collagen, elastin and their derivatives, wherein said method comprises transformation of the chloroplast genome of microalgae with a nucleic acid sequence encoding said recombinant protein, polypeptide or peptide.
Regarding claim 17, claim 3 of App’560 recites a method according to claim 2, comprising: (i) providing a nucleic acid sequence encoding said recombinant protein, polypeptide or peptide; (ii) introducing the nucleic acid sequence according to (i) into an expression vector which is capable of expressing the nucleic acid sequence; and (iii) transforming the chloroplast genome of microalgae host cell by the expression vector.
Regarding claim 18, claim 4 of App’560 recites a method according to claim 3, further comprising: (iv) identifying the transformed microalgae host cell; (v) characterizing the microalgae host cell for the production of recombinant protein, polypeptide or peptide expressed from said nucleic acid sequence; (vi) extracting the recombinant protein, polypeptide or peptide; and optionally (vii) purifying the recombinant protein, polypeptide or peptide.
Regarding claim 28 and 35, claim 11 of App’560 recites a method according to claim 3, wherein said at least one expression cassette further comprises a nucleic acid sequence encoding a signal peptide.
Regarding claims 24-25 and 32, claim 13 of App’560 recites a method according to claim 2, wherein said microalgae is chosen from the group consisting of Chlorophyta, Chlorophyceae, Pleurastrophyceae, Prasinophyceae, Chromophyta, Bacillariophyceae (diatoms), Chrysophyceae, Phaeophyceae, Eustigmatophyceae, Haptophyceae, Raphidophyceae, Xanthophyceae, Cryptophyta, Cryptophyceae, Rhodophyta, Porphyridiophycea, Stramenopiles, Glaucophyta, Glaucocystophyceae, Chlorarachniophyceae, Haptophyceae, Dinophyceae, Scenedesmaceae, Euglenophyta, Euglenophyceae. Claim 14 of App’560 recites a method according to claim 13, wherein said microalgae is chosen from the group consisting of Chlamydomonas, Chlorella, Dunaliella, Haematococcus, diatoms, Scenedesmaceae, Tetraselmis, Ostreococcus, Porphyridium, and Nannochloropsis.
The copending claims do not teach wherein the recombinant protein comprises aprotinin or that the fusion protein comprises a signal peptide.
Paget teaches a method of expressing a recombinant chimeric protein of interest in the thylakoid lumen of plants comprising transforming the cells with a vector encoding a bacterial signal peptide fused with a heterologous nucleic acid sequence encoding a recombinant or protein or peptide of interest (Abstract; p. 3, ln 22-25, p. 4, ln 1-8; p. 11, ln 19-22; p. 13, ln 31 – 33; p. 14, ln 8-15; p. 17, ln 31 – p. 18, ln 18; Claims 1-2, 5, 7-9, and 13-14). Paget teaches the fusion protein comprising a bacterial peptide is able to translocate across the thylakoid membrane and localize to the lumen of thylakoids in plants (p. 22, ln 26 – p. 24, ln 15). The transplastomic plant and/or progeny can comprise algae (p. 13, ln 31-33). The method leads to high-level accumulation of active and correctly folded recombinant enzyme in the thylakoid lumen of chloroplasts (Abstract; p. 23, ln 26- p. 24, ln 15). The thylakoid lumen of chloroplasts is a cellular compartment optimal for the accumulation of certain recombinant proteins due to its content of proteases (p. 2, ln 25-27). The peptide of interest can be aprotinin (p. 2, ln 31 – p. 3, ln 6; p. 11, ln 5-16; p. 15, ln 35 - p. 16, ln 2). Aprotinin is known to inhibit serine proteases including trypsin, chymotrypsin, plasmin and kallikrein (p. 11, ln 6-9). Paget teaches constructing a transformation vector by cloning a nucleic acid sequence of interest into an expression vector (p. 17, ln 31 – p. 18, ln 19).
Collier teaches a fusion protein comprising a protease inhibitor and a peptide of interest (Claim 1). In one embodiment, the fusion protein consists in order of a signal sequence, a peptide of interest, a cleavable linker region, and a protease inhibitor or variant thereof (para [0016]).
Itzhaki teaches proteolytic degradation of proteins in the thylakoid lumen, but discloses that the proteases involved are largely unknown (Abstract; p. 7094, col 2, para 2). Apolastocyanin is an example of a protein transported across the thylakoid membrane of in the thylakoid lumen of Chlamydomonas cells but it fails to accumulate due to rapid degradation (p. 7094, col 2, para 2). Serine protease DegP is implicated in the proteolytic degradation of proteins in the thylakoid lumen (Abstract). Isolated thylakoid membranes containing the chloroplast homologue of DegP degraded b-casein, an in vitro substrate of the bacterial protease; this activity was partially inhibited by a serine protease inhibitor, suggesting that at least part of the casein-degrading activity in the thylakoid membrane is attributable to DegP (Abstract). Based on these teachings, an ordinary artisan would immediately recognize that the thylakoid lumen of chloroplasts contains at least one serine protease that degrade proteins as well as the activity of the serine protease can be inhibited by serine protease inhibitors.
Stutz teaches the production of algae extracts and its use in cosmetics and pharmaceuticals to guard against extrinsic aging of the skin (Abstract). Application of extract inhibits the expression of matrix metalloproteinases 1 and 3, and matrix metalloproteinases 1 and 3 are higher in aged skin and responsible for the depletion of structure proteins like elastin and which play a role in skin aging (para [0026]).
Gunin teaches the total number of fibroblasts in skin decreases progressively with age and that the reduced number of fibroblasts is partly due to the reduced activity of proliferation in the cells (Abstract).
Koria teaches elastin-like polypeptides (termed ELPs) increase fibroblast proliferation (p. 1034, col 2, para 2). Koria teaches a fusion protein comprising ELPs (Abstract).
In view of the combination of claims 2-4, 11, and 13-14 of App’560 and the teachings Paget, Collier, Itzhaki, Stutz, Gunin, and Koria, it would have been obvious for a person of ordinary skill in the art that to modify the recombinant microalgae and method for producing a fusion protein comprising a signal peptide and a protein, polypeptide or peptide of interest, chosen from collagen, elastin and their derivatives, and wherein said algae and method comprises transformation of the chloroplast genome of microalgae with a nucleic acid sequence encoding said recombinant protein, polypeptide or peptide recited by App’560 such the signal peptide comprises the signal peptide taught by Paget that target the thylakoid lumen of chloroplasts, that peptide of interest is the ELP taught by Koria and is fused to a protease inhibitor as taught by Collier, and the protease inhibitor comprises aprotinin taught by Paget in order to increase the accumulation of the fusion protein or peptide of interest in the thylakoid lumen of chloroplasts, thereby arriving at the invention of claims 16-25, 28, 30-32, and 35-37.
An ordinary artisan would have been motivated to modify the fusion protein taught by Collier comprising a protease inhibitor and a peptide of interest such that the protease inhibitor comprises aprotinin taught by Paget because a simple substitution of the protease inhibitor for aprotinin would have been expected to yield predictable and expected results since Paget taught aprotinin is a protease inhibitor. An ordinary artisan would have been motivated to modify the nucleic acid encoding the fusion of a signal peptide to a protein or peptide of interest recited by App’560 such that the protein or peptide of interest is fused to a protease inhibitor as taught by Collier such as the serine protease aprotinin taught by Paget - in order to decrease proteolytic degradation of the chimeric protein and/or the protein of interest and therefore increase the accumulation of the chimeric protein and/or the protein of interest in the thylakoid lumen of chloroplasts. This is because Itzhaki taught the proteolytic degradation of proteins in the thylakoid lumen and implicated the role of serine proteinase DegP which can be partially inhibited by a serine protease inhibitor, and Paget taught aprotinin is a serine protease inhibitor. An ordinary artisan would have been motivated to substitute the protein of interest that is fused to aprotinin for the ELP taught by Koria so that the extract of the algae comprising the aprotinin-ELP fusion can be used in cosmetics or pharmaceutics to treat aging because Stutz taught the production of algae extracts and its use in cosmetics and pharmaceuticals to guard against aging of skin, Gunin taught the total number of fibroblasts in skin decreases progressively with age and that the reduced number of fibroblasts is partly due to the reduced activity of proliferation in the cells, and Koria teaches ELPs that increase the proliferation of fibroblasts. Therefore, the fusion of aprotinin to the protein of interest (i.e., ELP) would provide the advantage of inhibiting serine proteases in the thylakoid lumen of chloroplasts and result in the increased accumulation of the chimeric protein and/or the protein of interest in the thylakoid lumen of chloroplasts.
An ordinary artisan would have had a reasonable expectation of success of modifying the fusion protein taught by Collier comprising a protease inhibitor and a peptide of interest such that the protease inhibitor comprises aprotinin taught by Paget because a simple substitution of the protease inhibitor for aprotinin would have been expected to yield predictable and expected results since Paget taught aprotinin is a protease inhibitor. An ordinary artisan would have been motivated to modify the nucleic acid encoding the fusion of a signal peptide to a protein or peptide of interest recited by App’560 such that the protein or peptide of interest is fused to a protease inhibitor as taught by Collier such as the serine protease aprotinin taught by Paget because Collier taught the fusion of aprotinin to a peptide of interest. An ordinary artisan would have had a reasonable expectation of success substituting the signal peptide in the fusion protein recited by App’560 since substituting one signal peptide for another is expected to yield predictable results. An ordinary artisan would have had a reasonable expectation of success that the modified fusion protein comprising a signal peptide, aprotinin, and a protein or peptide of interest would decrease proteolytic degradation of the chimeric protein and/or the protein of interest and therefore increase the accumulation of the chimeric protein and/or the protein of interest in the thylakoid lumen of chloroplasts. This is because Itzhaki taught the proteolytic degradation of proteins in the thylakoid lumen and implicated the role of serine proteinase DegP which can be partially inhibited by a serine protease inhibitor, and Paget taught aprotinin is a serine protease inhibitor. An ordinary artisan would have been motivated to substitute the protein of interest that is fused to aprotinin for the ELP taught by Koria because Koria taught a fusion protein comprising ELP. Therefore, the fusion of aprotinin to the protein of interest (i.e., ELP) would provide the advantage of inhibiting serine proteases in the thylakoid lumen of chloroplasts and result in the increased accumulation of the chimeric protein and/or the protein of interest in the thylakoid lumen of chloroplasts.
For the reasons stated herein, claims 16-25, 28, 30-32, and 35-37 of this application are unpatentable over claims 2-4, 11, and 13-14 of App’560 in view of Paget, Collier, and Itzhak. This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
The provisional rejection of claims 26, 29, and 33 on the ground of nonstatutory double patenting as being unpatentable over claims 2-4, 11, and 13-14 of copending Application No. 17/802,560 App’560” in view of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria as applied to claims 16-25, 28, 30-32, and 35-37 above, and further in view of Chen is maintained.
The combination of claims 2-4, 11, and 13-14 of App’560 and the teachings of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria, as applied to claims 16-25, 28, 30-32, and 35-37, are discussed above and incorporated herein.
Regarding claim 26 and 33, the combination of claims 2-4, 11, and 13-14 of App’560 and the teachings of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria does not explicitly teach wherein the recombinant protein, polypeptide or peptide of interest is fused to the C-terminus of aprotinin.
Chen teaches creating fusion proteins comprising in all possible directions (i.e., (Protein A)-(Protein B) and (Protein B)-(Protein A)) in order to test the optimal orientation (p. 1363, col 2, para 3).
In view of the combination of claims 2-4, 11, and 13-14 of App’560 and the teachings of Paget, Collier, Itzhaki, Stutz, Gunin, Koria, and Chen, it would have been obvious for a person of ordinary skill in the art that the aprotinin fusion protein taught by the combination of App’560, Paget, Collier, Itzhaki, Stutz, Gunin, Koria comprise the protein, polypeptide, or peptide of interest (i.e., collagen or elastin) fused to the C-terminus of aprotinin, thereby arriving at the invention of claim 26 and 33.
An ordinary artisan would have been motivated to fuse the protein, polypeptide, or peptide of interest the C-terminus of aprotinin in order to determine the most effective orientation of the fusion protein since Chen taught producing fusion proteins in a plurality of orders in to determine the optimal orientation of the peptides within the fusion protein.
An ordinary artisan would have had a reasonable expectation of success of fusing a protein, polypeptide, or peptide of interest is fused to the C-terminus of aprotinin since Chen taught producing fusion proteins in a plurality of orders in to determine the optimal orientation of the peptides within the fusion protein.
Regarding claim 29, Paget further teaches the production of a chimeric protein comprising a peptide of interest in a transplastomic plant cell wherein the nucleic acid encoding the protein of interest is fused to a bacterial signal peptide and the signal peptide is cleaved after expression (Abstract; p. 13, ln 19-29; p. 14, ln 8-17; p. 15, ln 4-28; p. 16, ln 14-25; Claims 1, 7-9, 12, 15).
The combination of claims 2-4, 11, and 13-14 of App’560 and the teachings of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria does not explicitly teach wherein the recombinant protein, polypeptide or peptide of interest and aprotinin wherein the fusion protein also comprises a protease specific cleavage site between the aprotinin and the recombinant protein, polypeptide or peptide of interest.
Chen teaches protease specific cleavable linkers between domains of fusion proteins (Abstract; p. 1361, col 2, para 3; Tables 2-3, Figure 2-4). Cleavable linkers enable targeting but also release a free functional domain of a fusion protein after cleavage (Abstract; Table 3).
In view of the combination of claims 2-4, 11, and 13-14 of App’560 and the teachings of Paget, Collier, Itzhaki, Stutz, Gunin, Koria, and Chen, it would have been obvious for a person of ordinary skill in the art before the effective filing date of the claimed invention a recombinant algae and method of producing a recombinant protein comprising a signal peptide, aprotinin, and collagen or elastin in algae taught by the combination of App’560, Paget, Collier, Itzhaki, Stutz, Gunin, and Koria to comprise between the aprotinin and the protein, polypeptide, or peptide of interest (i.e., collagen or elastin) a protease specific cleavable linker taught by Chen, thereby arriving at the invention of claim 29.
An ordinary artisan would have been motivated that the recombinant protein comprising a signal peptide, aprotinin, and collagen or elastin comprise between the aprotinin and the protein, polypeptide, or peptide of interest is a protease specific cleavable linker taught by Chen in order to free the protein or peptide of interest from the fusion protein for isolation. This is because Collier taught a fusion protein comprising a cleavable linker between a protein or peptide of interest and a protease inhibitor and Chen taught protease specific cleavable linkers as well as that cleavable linkers allow for the release a free functional domain of a fusion protein after cleavage.
An ordinary artisan would have had a reasonable expectation of success of modifying the aprotinin fusion protein taught by Paget to comprise between the aprotinin and the protein, polypeptide, or peptide of interest a protease specific cleavable linker taught by Chen because Chen teaches the use of protease specific cleavable linkers between domains of fusion proteins.
For the reasons stated herein, claims 26, 29, and 33 of this application are unpatentable over claims 2-4, 11, and 13-14 of App’560 in view of Paget, Collier, Itzhak, and Chen. This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
The provisional rejection of claims 27 and 34 on the ground of nonstatutory double patenting as being unpatentable over claim 2-4, 11, and 13-14 of copending Application No. 17/802,560 App’560” in view of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria as applied to claims 16-25, 28, 30-32, and 35-37 above, and further in view of Mayfield is maintained.
The combination of claims 2-4, 11, and 13-14 of App’560 and the teachings of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria, as applied to claims 16-25, 28, 30-32, and 35-37, are discussed above and incorporated herein.
Regarding claims 27 and 34, Claim 10 of App’560 further limits the method of claim 3, wherein said at least one expression cassette further comprises a nucleic acid sequence encoding an epitope tag peptide fused operationally at its 5' or 3' end to the said nucleic acid sequence encoding the recombinant protein, polypeptide or peptide.
The combination of claims 2-4, 11, and 13-14 of App’560 and the teachings of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria does not explicitly teach wherein the recombinant protein, polypeptide or peptide of interest is comprises an epitope tag.
Mayfield teaches the expression of fusion protein in algae comprising a FLAG epitope tag for the purpose of analyzing the expression of the protein as well as for using affinity purification (Title; Abstract; para [0155, 0174]; Claim 21). The algae can comprise Chlamydomonas reinhardtii (para [0032]).
In view of the combination of claims 2-4, 11, and 13-14 of App’560 and the teachings of Paget, Collier, Itzhaki, Stutz, Gunin, and Mayfield, it would have been obvious for a person of ordinary skill in the art to have modified the recombinant algae and modified method for producing a recombinant protein in algae comprising a signal peptide, aprotinin, and polypeptide or peptide comprising wherein said protein, polypeptide or peptide is chosen from collagen or elastin taught by the combination of App’560, Paget, Collier, Itzhaki, Stutz, Gunin, and Koria such that the recombinant protein comprises an epitope tag, thereby meeting the limitations of claims 27 and 34 of the instant application.
An ordinary artisan would have been motivated to have modified the recombinant algae and modified method for producing a recombinant protein in algae comprising a signal peptide, aprotinin, and polypeptide or peptide comprising wherein said protein, polypeptide or peptide is chosen from collagen or elastin taught by the combination of App’560, Paget, Collier, Itzhaki, Stutz, Gunin, and Koria such that the recombinant protein comprises an epitope tag in order to analyze the protein and to isolate it via affinity purification since the App’560 taught an epitope tag to the recombinant protein or polypeptide and Mayfield taught the use of a FLAG epitope tag in a fusion protein produced in algae for the purpose of analyzing the expression of the protein as well as for using affinity purification.
An ordinary artisan would have had a reasonable expectation of success of modifying the recombinant algae and modified method for producing a recombinant protein in algae comprising a signal peptide, aprotinin, and polypeptide or peptide comprising wherein said protein, polypeptide or peptide is chosen from collagen or elastin taught by the combination of App’560, Paget, Collier, Itzhaki, Stutz, Gunin, and Koria such that the recombinant protein comprises an epitope tag since the App’560 recites an epitope tag to the recombinant protein or polypeptide and Mayfield taught the use of a FLAG epitope tag in a fusion protein produced in algae for the purpose of analyzing the expression of the protein as well as for using affinity purification.
For the reasons stated herein, claims 27 and 34 of this application are unpatentable over claims 2-4, 11, and 13-14 of App’560 in view of Paget, Collier, Itzhaki, Stutz, Gunin, and Koria, and Mayfield. This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
RESPONSE TO REMARKS: Applicant submits that the instant claims are not disclosed or suggested by copending Application No. 17/802,560 and the cited prior art, taken either singly or in combination as well as that the copending Application No. 17/802,560 differs from the present claims.
Applicant’s arguments are not found persuasive. For the reasons set forth in the new provisional rejections above, it would have been obvious to one of ordinary skill in the art to modify the co-pending claimed drawn to a method for producing a recombinant protein, polypeptide or peptide comprising repeat units of amino acids in the chloroplast of microalgae, said protein, polypeptide or peptide being chosen from collagen, elastin and their derivatives, wherein said method comprises transformation of the chloroplast genome of microalgae with a nucleic acid sequence encoding said recombinant protein, polypeptide or peptide of App’560 such that that method comprises a fusion protein aprotinin with an ELP in order to increase the accumulation of the ELP and/or the fusion protein comprising thereof as well as meets all of the claim limitations of the instant claims.
Conclusion
No claims are currently allowed for the reasons as stated above. Applicants must
respond to the objections/rejections in this Office action to be fully responsive in
prosecution.
The instant Office Action is non-final.
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/SCOTT E. MULDER/Examiner, Art Unit 1656
/David Steadman/Primary Examiner, Art Unit 1656