DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Election/Restrictions
Applicant’s election without traverse of Group I (claims 1-5,8-17 and 21-24) and species of full-length human GLUT1, SEQ ID NOS: 1 (LALBA) and 2 (B4GalT1) sialyl-lactose and
PNG
media_image1.png
165
643
media_image1.png
Greyscale
in the reply filed on 06/01/2026 is acknowledged.
Claim 18 is withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected Invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 06/01/2026.
Claim 11 is withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected species, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 06/01/2026. Elected SEQ ID NO: 1 and 2 are human LALBA and B4GalT1; as such claim 11 does not read on the elected species.
Claim Objections
Claims 14 and 15 are objected to because of the following informalities:
Claim 14, recites “the sequences encoding alpha-lactalbumin (LALBA), beta-1,4-galactosyltransferase 1 (B4GaIT1), glucose transporter, and combinations thereof.” Claim 3 from which claim 14 depends requires sequences encoding all of alpha-lactalbumin (LALBA), beta-1,4-galactosyltransferase 1 (B4GaIT1), and glucose transporter be encoded on at least one expression vector. As such, recitation of “combinations thereof” is superfluous since a combination is required by any embodiment of claims 3 and 14. See rejection under 35 U.S.C. 112(d) below.
Claim 15 is a length claim of Markush group species; as such, only the first species is directly addressed. Claim 15 should be modified to indicate:
-[[an]] a first expression construct that encodes an alpha-lactalbumin (LALBA), a beta-1,4- galactosyltransferase 1 (B4GaIT1), a glucose transporter 1 (GLUT1), and a marker protein, wherein C-terminus of LALBA is linked to N-terminus of B4GaIT1, C-terminus of B4GaT1 is linked to N-terminus of GLUT1, and C-terminus of GLUT1 is fused to N-terminus of the marker protein;
and [[an]] a second expression construct that encodes a beta-1,4-galactosyltransferase 1 (B4GaIT1) and an alpha-lactalbumin (LALBA), wherein C-terminus of B4GaT1 is linked to N-terminus of LALBA.
That is, some punctation, such as a semicolon or comma, should be present between the two paragraphs and that two separate expression constructs are being defined should be highlight by labeling the expression constructs as first and second, or similar language indicating two expression constructs are required. Many of the Markush species recited in claim 15 have parallel formatting and should be similarly corrected.
Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 16 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 16 recites “wherein the expression constructs comprise, if present, the following.” The recitation of “if present” raises substantial doubt as to whether the claim features that follow (identity to specific sequence identifiers) are or are not limitations of the claimed invention. That is, recitation of “if present” raises the possibility that “not present” is an embodiment of claim 16. However, if the remaining features of claim 16 are “optional,” then claim 16 would not further limit claim 1 as required by 35 U.S.C. 112(d). Since it is not clear as to whether the claim features after “following” are or are not limitations of the claimed invention, an ordinarily skilled artisan at time of filing cannot determine how to avoid infringement of claim 16.
The following is a quotation of 35 U.S.C. 112(d):
(d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph:
Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
Claim 14 is rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends.
Claim 14 depends from claim 3 that in turn depends from independent claim 2. Claim 3 requires “one or more expression constructs” encoding LALBA, B4GalT1 and an additional glucose transporter. Recitation in claim 14 of “at least one, at least two, at least three or at least four constructs” is a synonym for “one or more expression constructs.” The only required featured of claim 14 is one expression construct where embodiments being two, three, four or more expression constructs are optional and not limiting features of claim 14. Since claim 3 from which claim 14 depends already requires LALBA, B4GalT1 and an additional glucose transporter be encoded by at least one (i.e. one or more) expression constructs, claim 14 does not further limit claim 3.
Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements.
Claim Rejections - 35 USC § 102
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.
(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.
Claim(s) 1, 2, 12, 13 and 24 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Do (Generation of novel chimeric LacdiNAcS by gene fusion of α-lactalbumin and β1,4-galactosyltransferase 1, Glyconj J 26, 2009, 567-75).
Do, abstract, states:
Novel chimeric lacdiNAc (GalNAc(β1-4)GlcNAc) synthase (c-LacdiNAcS) was generated by gene fusion of α-lactalbumin (α-LA) and β1,4-galactosyltransferase 1 (β1,4-GalT1). c-LacdiNAcS was expressed in Lec8 Chinese hamster ovary (Lec8 CHO) cells and exhibited N-acetylgalactosaminyltransferase (GalNAcT) activity in the absence of exogenous α-LA as well as other glycosyltransferase activities including lactose synthase (LacS), and β1,4-GalT. These glycosyltransferase activities of c-LacdiNAcS were compared to those activities induced in LacS system under the co-presence of bovine β1,4-GalT1 and α-LA, indicating that each domain of α-LA and β1,4-GalT1 on c-LacdiNAcS is not only folding correctly, but also interacting together. Furthermore, c-LacdiNAcS was found to be auto-lacdiNAcylated and can synthesize lacdiNAc structures on cellular glycoproteins, demonstrating that GalNAcT activity of c-LacdiNAcS is functional in Lec8 CHO cells.
“In lactating mammary gland where high concentration of α-LA is present, β1,4-GalT1 changes its acceptor substrate specificity from N-acetylglucosamine (GlcNAc) to glucose and subsequently, produces lactose through the induced lactose synthase (LacS) activity.” Do, page 567, right col.
“Lec8 Chinese hamster ovary (Lec8 CHO cells, ATCC) were transfected with the chimeric fusion construct (pcDNA6-V5-His/α-LA:β1,4-GalT1) using LipofectAmineTM (Invitrogen) according to manufacturer’s instructions. Stable cells were screened under 10 μg/ml blasticidin.” Do, section 1.2
The chimeric fusion construct (pcDNA6-V5-His/α-LA:β1,4-GalT1) described is understood to be a mammalian expression construct comprising polynucleotides encoding LALBA and B4GalT1 that is comprised in a recombinant cell as recited in claims 2, 12 and 24.
Regarding claim 1, the described fusion of α-lactalbumin (α-LA) and β1,4-galactosyltransferase 1 (β1,4-GalT1) is described to have lactose synthase activity as shown in Fig. 3B of Do. The preamble of claim 1 of a “recombinant cell for producing lactose” is a statement of intended use. “If the body of a claim fully and intrinsically sets forth all of the limitations of the claimed invention, and the preamble merely states, for example, the purpose or intended use of the invention, rather than any distinct definition of any of the claimed invention’s limitations, then the preamble is not considered a limitation and is of no significance to claim construction.” MPEP 2111.02(II). Claim 1 does not require lactose to be produced, but only that an embodiment recombinant cell have some latent ability for lactose production. Since the described fusion of α-lactalbumin (α-LA) and β1,4-galactosyltransferase 1 (β1,4-GalT1) as expressed in a CHO cell has lactose synthase ability, the features of claims 1 and 13 are disclosed by Do.
Claim Rejections - 35 USC § 103
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.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claim(s) 1-5, 8-10, 12-15 and 21-24 is/are rejected under 35 U.S.C. 103 as being unpatentable over Rye et al. (WO 2021/067641 A1) (see IDS, 05/16/26) further in view of Do (Generation of novel chimeric LacdiNAcS by gene fusion of α-lactalbumin and β1,4-galactosyltransferase 1, Glyconj J 26, 2009, 567-75).
Rye, abstract, states:
Compositions, kits, and methods for preparing nutrient compositions are provided. A nutrient composition can be a food composition such as a dairy composition. A nutrient composition may be produced by an engineered cell composition. Compositions, kits, and methods for preparing engineered cell compositions for producing nutrient compositions are also provided. An engineered cell composition may comprise genetically engineered cells, such as genetically engineered mammary or mammary-like cells. An engineered cell composition can be derived from mammalian stem cells, such as non-mammary adult stem cells. Related methods of characterization, such as for characterizing the engineered cell compositions or the nutrient compositions, are also provided.
“In some embodiments of the method for generating the engineered cell, the modified expression profile comprises an up-regulated expression of a gene or a transcript that regulates production of a milk protein. In some embodiments, the modified expression profile comprises a down-regulated expression of a gene or a transcript that regulates production of a milk protein. In some embodiments, the gene or the transcript regulates production of a milk protein selected from the group consisting of: a S1/S2 casein, b casein, k casein, a-lactalbumin, lactoferrin, lactoperoxidase, lysozyme, haptocorrin, osteopontin, serum albumin, complement C3, complement C4, and a combination thereof.” Rye, para. [0018].
“The stem cells, mammary cells, or a mammary-like cells can be engineered such that the genome of the engineered stem cells, engineered mammary cells, or engineered mammary-like cells is altered, or different, from the genome of the respective naturally occurring cells. Such engineered cells can have genes upregulated to increase cell proliferation. Such engineered cells can have their sensitivity to different growth factors increased or enhanced by upregulating a corresponding receptor in the cell. Upregulation can be achieved through various known method(s) or mechanism(s), including those described herein, and can include epigenetic remodeling, changing of regulatory sequences, removing regulatory molecules, substitution of promoter elements, increasing copy number of genes, or turning genes constitutively on or off.” Rye, para. [00228].
“Upregulation of lactose synthesis can be done through the upregulation of at least one of AKT1, GLUT1 and lactose synthase (a-lactalbumin and b1,4- galactosyltransferase) genes to increase glucose influx and conversion of glucose to lactose within the cells.” Rye, para. [00229]. “Upregulation can be achieved through various known method(s) or mechanism(s), including those described herein, and can include epigenetic remodeling, changing of regulatory sequences, removing regulatory molecules, substitution of promoter elements, increasing copy number of genes, or turning genes constitutively on or off.” Rye, para. [00228].
“The raising or increasing of a concentration or amount of a milk component in a liquid composition may be desired. For example, the gene, genetic construct, or expression mechanism, for a certain lipid or fat can be upregulated, in a mammary cell, or mammary-like cell. The gene, genetic construct, or expression mechanism, for a certain protein can be upregulated in a mammary cell, or a mammary-like cell. For example, the gene, genetic construct, or expression mechanism, for a certain oligosaccharide can be upregulated, in a mammary cell, or a mammary-like cell. In another example, human mammary cells or human mammary-like cells can be engineered or modulated to upregulate Secretor gene that encodes fucosyltransferase-2 (FUT-2) or the Lewis gene that encodes fucosyltransferase-3 (FUT-3) enzyme expression levels. These genes may be involved in the production of Human Milk Oligosaccharides (HMOs). An upregulation may comprise a gene knock in. As used herein, the term “knock-in” generally refers to the insertion of an exogenous or a heterologous gene into a specific locus in the genome of a cell. The knocked-in exogenous gene can be used to analyze product function and/or expression pattern. The exogenous gene can be introduced to disrupt the endogenous gene, generating a knock-out/knock-in cell. The knock-in gene can be used with an inducible gene expression strategy with, for example, a Cre/loxP system, a ubiquitous gene expression strategy using ROSA26 locus, or other various strategies.” Rye, para. [00202].
“This example illustrates a method for CRISPR/Cas9 engineering of mammary or mammary like cells as described in the present disclosure for knocking in an exogenous nucleic acid. In some cases, the exogenous nucleic acid can be a regulatory region such as enhancer, promoter, silencer, insulator, or operator. In some cases, the exogenous nucleic acid comprises a promoter, where the promoter can be an inducible promoter (e.g. the inducible promoter system described in FIG. 4). In some cases, the exogenous nucleic acid being knocked in the mammary or mammary-like cells by the CRISPR/Cas described herein can encode any one of the gene described herein. In some cases, the exogenous nucleic acid can encode an exogenous gene.” Rye para. [00318]. “At least one vector encoding the components of the CRISPR/Cas9 (e.g. gRNA, exogenous nucleic acid to be knocked in, or Cas9) is introduced into the mammary or mammary-like cell. Cas9 performs a site-directed digestion to cut the chromosome of the mammary or mammary-like cell at a precise location targeted by the gRNA. The break in the chromosome is subsequently repaired by inserting the exogenous nucleic acid into the chromosome, thus knocking in the exogenous nucleic acid comprising the exogenous regulatory region or the exogenous gene.” Rye, para. [00319].
“An engineering gene vector may be developed from the target gene identification using a specific choice of vector, insertion of the target gene, and choice of promotor, and upon insertion into a cell, genetic manipulation may be acquired wherein gene expression may be up- or down-regulated.” Rye, para. [00198].
To summarize the above, Rye directly suggests in a mammary or mammary-like cell or mammalian stem cell that “Upregulation of lactose synthesis can be done through the upregulation of at least one of AKT1, GLUT1 and lactose synthase (a-lactalbumin and b1,4- galactosyltransferase) genes to increase glucose influx and conversion of glucose to lactose within the cells.” Rye, para. [00229]. As such, at the time of filing an ordinarily skilled artisan would have been motivated to following these express teachings of Rye in order to achieve the benefit of a mammary or mammary-like cell or mammalian stem cell having increased glucose influx and conversion of glucose to lactose by upregulation of at least GLUT1 and lactose synthase comprising alpha-lactalbumin (LALBA) and beta-1,4-galactosyltransferase. Rye does not expressly states that the beta-1,4-galactosyltransferase to be upregulated is specifically B4GalT1 as recited. However, Do, page 567, right col., explains that it is well-understood in the art that: “In lactating mammary gland where high concentration of α-LA is present, β1,4-GalT1 changes its acceptor substrate specificity from N-acetylglucosamine (GlcNAc) to glucose and subsequently, produces lactose through the induced lactose synthase (LacS) activity. Recent animal studies in vivo show that β1,4-GalT1 is indispensable to produce lactose in mammary gland. Among mammalian β1,4-GalTs, β1,4-GalT1 shows to induce LacS activity in presence of α-LA.” Do, page 567, right col. As such, at the time of filing in view of the teachings of Do, an ordinarily skilled artisan at time of filing would have understood that the specific species of b1,4-galactosyltransferase to be combined with alpha-lactalbumin for lactose synthesis is β1,4-GalT1 as recited in claim 1.
Again, at the time of filing an ordinarily skilled artisan would have been motivated to following these express teachings of Rye in order to achieve the benefit of a mammary or mammary-like cell or mammalian stem cell having increased glucose influx and conversion of glucose to lactose by upregulation of at least GLUT1 and lactose synthase comprising alpha-lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1). Rye states several methods for upregulation including increasing copy number of genes. An ordinarily skilled artisan at time of filing would have understood that increasing copy number of genes requires insertion of additional copies of the genes in question into the cell. That is, increasing copy number of a gene is done by insertion of additional or exogenous genes (nucleic acids) into the mammalian cell to be engineered. Rye teaches that addition or knock-in of additional exogenous genes into a mammalian cell can be done with a CRISPR-based method wherein “At least one vector encoding the components of the CRISPR/Cas9 (e.g. gRNA, exogenous nucleic acid to be knocked in, or Cas9) is introduced into the mammary or mammary-like cell.” Rye, para. [00319]. As such, at the time of filing in view of the teachings of Rye and Do, an ordinarily skilled artisan at the time of filing would have been motivated to produce one or more vectors encoding GLUT1 (glucose transporter) and lactose synthase comprising alpha-lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1) suitable for introduction into mammalian cells as described by Rye to allow for the same cells to produce or increase production of lactose, whether by a CRISPR-based method as described by Rye or otherwise. As far as the gene encoding GLUT1 (glucose transporter) and lactose synthase comprising alpha-lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1) are intended to be expressed in such mammalian cells, any vector accomplishing such expression is within the broadest reasonable meaning of an “expression construct” or “mammalian expression vector.” GLUT1 as taught by Rye is understood to refer to a complete or full-length GLUT1. For these reasons, the combined teachings of Rye and Do teach the features of claims 1-5, 8, 12-14 and 24.
Regarding claims 9 and 10, as discussed, Rye teaches upregulation of genes encoding GLUT1 (glucose transporter) and lactose synthase comprising alpha-lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1). The same directly suggests that the same genes subject to upregulation by increase in copy number or otherwise are otherwise endogenous in the mammalian cell to be modified. “In some embodiments, the mammary epithelial cells are of human origin.” Rye et al., para. [0006]. “Fresh adipose tissue is obtained from slaughterhouses or liposuction (e.g., in case of humans, after informed consent), kept in phosphate buffer saline supplemented with antibiotics.” Rye, para. [00297]. Since Rye directly suggests engineered human cells, an ordinarily skilled artisan at the time of filing in upregulation of genes encoding GLUT1 (glucose transporter) and lactose synthase comprising alpha-lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1), as discussed above, by insertion of additional copies of such genes through an expression construct would have been motivated to choose or select human sourced genes (all from the same mammalian species) in order to affect upregulation as discussed.
Regarding claim 15, claim 15 recites a Markush group that includes the following:
PNG
media_image2.png
153
654
media_image2.png
Greyscale
“As used herein, the term “knock-in” generally refers to the insertion of an exogenous or a heterologous gene into a specific locus in the genome of a cell.” Rye, para. [00202]. “At least one vector encoding the components of the CRISPR/Cas9 (e.g. gRNA, exogenous nucleic acid to be knocked in, or Cas9) is introduced into the mammary or mammary-like cell. Cas9 performs a site-directed digestion to cut the chromosome of the mammary or mammary-like cell at a precise location targeted by the gRNA. The break in the chromosome is subsequently repaired by inserting the exogenous nucleic acid into the chromosome, thus knocking in the exogenous nucleic acid comprising the exogenous regulatory region or the exogenous gene.” Rye, para. [00319].
Although alpha-lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1) can act together in one complex as a lactose synthase, the same are separate genes and in Rye are not taught to be present in a fusion protein. As far as it is a valid design choice for copies the two independent alpha-lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1) genes to be expressed from different genomic loci (e.g. inserted into different chromosomes or different regions of the same chromosome), the same would require the same to be present in different vectors or expression constructs encoding gRNA as described targeting such different genetic loci. That is, the arrangement of parts of genes encoding lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1) into the same or different expression constructs (i.e. two constructs, a first construct and a second construct) is a matter of rearrangement of parts or obvious matter of design choice in the art particularly in view of a possibility to target the two genes to different genetic loci. See MPEP 2144.04(VI)(C).
Regarding claims 21-23, “In some embodiments of the method for generating the engineered cell, the modified expression profile comprises an up-regulated expression of a gene or a transcript that regulates production of a milk carbohydrate. In some embodiments, the modified expression profile comprises a down-regulated expression of gene or a transcript that regulates production of a milk carbohydrate. In some embodiments, the gene or the transcript regulates production of a milk carbohydrate selected from the group consisting of: 2’-fucosyllactose, 3-fucosyllactose, difucosyllactose, difucosyllacto-N-tetrose (DFLNT), difucosyllacto-N-hexaose, fucosyllacto-N-hexaose (FLNH), lacto-N-fucopentaose (LNFP) I, LNFP II, LNFP III, 3’-sialyllactose, 6’-sialyllactose, disialyllacto-N-hexaose (DSLNH), disialyllacto-N-tetraose (DSLNT), fucodisialyllacto-N-hexaose (FDSLNH), sialyl-lacto-N-tetraose b (LSTb), and sialyl-lacto-N- tetraose c (LSTc), lacto-N-hexaose, lacto-N-neotetraose (LNnT), lacto-N-tetrose (LNT), lactose, and a combination thereof. In some embodiments, the gene or the transcript regulating the production of the milk carbohydrate is a gene or a transcript selected from the group consisting of: LALBA, galactosyltransferase B4GALT1, galactosyltransferase B4GALT2, LGALS1, LGALS2, LGALS3, LGALS8, LGALS9, LGALS12, LGALS13, LGALS14, LGALS16, LCT, GLUT1, FUT1, FUT2, FUT3, FUT4, FUT5, FUT6, FUT7, FUT8, FUT9, sialyltransferase, GALM, GLTP, LGALSL, N-acetylglucosaminyltransferase, glycosyltransferase, and a combination thereof.” Rye, para. [0019]. “In some embodiments, the oligosaccharide comprises from 3 to 20 saccharide units. In some embodiments, the oligosaccharide comprises a terminal lactose moiety.” Rye, para. [0061]. “Each of the HMOs structure may start with a lactose unit “Gal (b1-4) Glc” which results from formation of a b 1-4 glycosidic linkage between galactose and glucose catalyzed by the lactose synthase protein complex. Several tri-saccharides can be synthesized by appending either galactose or fucose to the reducing or non-reducing end of the lactose residue, which is performed through galactosyl- or fucosyl-transferase activity. Resulting components may be for example, lacto-N-neoetraose (LNnT), 3'-galactosyllactose (Gal^l-3)Gal^l-4)Glc), 4'-galactosyllactose (Gal^l- 4)Gal^l-4)Glc), 6'-galactosyllactose (Gal^l-6)Gal^l-4)Glc), 2'-FL (Fuc(a l-2)Gal^l-4)Glc), and 3- fucosyllactose (3’-FL) (Oh1(b 1 -4) [ Fuca 1 -3 |Glc). If sialic acids are connected to the non-reducing end of lactose via sialyl-transferases, 3'-sialyllactose (3'-SL; Neu5Ac (a 2-3) Gal^l-4) Glc) and 6'-sialyllactose (6'-SL) (Neu5Ac (a 2-6) Oh1(b 1 -4) Glc) are formed.” Rye, para. [00119].
That is, a purpose of upregulation of LALBA and B4GalT1 for production of lactose, as discussed above, is to in part serve as a basis for the synthesis of larger oligosaccharides. As explained by Rye, para. [00119], a sialyl-transferase can be further expressed in the engineered mammalian cells described in order to convert lactose to 3’-sialylactose such that an ordinary skilled artisan at time of filing would have been motivated to do the same as expressly taught by Rye to meet the features of claims 21-23.
Claim(s) 1-5, 8-10, 12-17 and 21-24 (all non-withdrawn claims) is/are rejected under 35 U.S.C. 103 as being unpatentable over Rye et al. (WO 2021/067641 A1) further in view of Do (Generation of novel chimeric LacdiNAcS by gene fusion of α-lactalbumin and β1,4-galactosyltransferase 1, Glyconj J 26, 2009, 567-75) as applied to claims 1-5, 8-10, 12-15 and 24 above, and further in view of GenBank, Accession No. BC069103.1, 2006, www.ncbi.nlm.nih.gov; GenBank, Accession No. AB590749.1, 2016, www.ncbi.nlm.nih.gov; Uniprot, Accession No. P11166, 2020, www.uniprot.org; and GenBank, Accession No. KJ892146.1, 2015, www.ncbi.nlm.nih.gov.
Regarding claims 16 and 17, as discussed in connection with claims 9 and 10 above, Rye suggests that in embodiments wherein an engineered cell is a human cell that the upregulated genes be GLUT1, lactalbumin (LALBA) and beta-1,4-galactosyltransferase I (β1,4-GalT1) genes from human.
GenBank BC069103.1 evidences that cDNA encoding for alpha-lactalbumin from humans (Homo sapiens) contains a sequence identical to recited SEQ ID NO: 1.
GenBank AB590749.1 evidences a DNA sequence encoding B4GalT1 from human contains a sequence identical to recited SEQ ID NO: 2.
Uniprot P11166 evidences the amino acid sequence for human GLUT1, which is also known as Glucose transporter type 1. GenBank KJ892146.1 evidences that the same protein is encoded by a DNA sequence containing recited SEQ ID NO: 3.
In implementing embodiments of Rye wherein a mammalian cell has increased copy number of alpha-lactalbumin, B4GalT1 and GLUT1 genes in order to affect upregulation by insertion of additional copies of these genes as discussed, the use of a specific encoding gene sequence is required. The cited GenBank and Uniprot references above indicate that SEQ ID NO: 1 is a known sequence that encodes LALBA, SEQ ID NO: 2 is a known sequence that encodes B4GalT1 and SEQ ID NO: 3 is a known sequence that encodes GLUT1 (all from human) such that an ordinarily skilled artisan at time of filing would have been motived to select and to have an expectation of success in utilizing such known sequence to introduce increased copy number of human LALBA, B4GalT1 and GLUT1 genes, since the same nucleotide sequences are expressly taught to encode the same proteins.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to TODD M EPSTEIN whose telephone number is (571)272-5141. The examiner can normally be reached Mon-Fri 9:00a-5:30p.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Robert Mondesi can be reached at (408) 918-7584. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/TODD M EPSTEIN/Primary Examiner, Art Unit 1652
Prosecution Insights