DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Claims 14, 16, 20-27, 29-35 are pending and under consideration in this Office Action.
In view of the claim amendment and arguments filed 01/02/2026, all previous claim objections and/or claim rejections have been withdrawn in favor of the instant claim rejection(s) stated below.
Claim Rejections - 35 USC § 112(b) or 35 U.S.C. 112 (pre-AIA ) 2nd Paragraph
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.
Claims 14, 16, 20-27, 29-31, 34, 35 are 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 pre-AIA the applicant regards as the invention.
Amended claims 14 encompasses any method for recovering a secreted compound of interest in a hairy roots- based expression system. However, the claim is vague and indefinite since specific methods steps for recovering the secreted compound of interest have not been recited. Dependent claims 16, 20-27, 29-31, 34, 35 are also rejected because they do not correct the defect.
Claim Rejections - 35 USC § 103
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 of this title, 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.
Claims 32, 33 are rejected under 35 U.S.C. 103 as being unpatentable over US20190040407 (02/07/2019; reference of record) in view of US20130061351 (03/07/2013; reference of record), Lallemand et al. (Extracellular peptidase hunting for improvement of protein production in plant cells and roots", FRONTIERS IN PLANT SCIENCE, vol. 6, 6 February 2015 (2015-02-06); IDS filed 05/24/2023).
According to MPEP 2111, while claims must be given their broadest reasonable interpretation consistent with the specification, limitations of the specification cannot be read into the claims to thereby narrow the scope of the claims. The claims have not been amended as claim 14 to specifically recite a solution comprising from about 250 mM to about 4 M of a salt selected from a group consisting of sodium salt, potassium salt, chloride salt, sulfate salt, nitrate salt, ammonium salt, phosphate salt, and combinations thereof or, from about 250 mM to about 4 M of a salt selected from the group consisting of sodium chloride, potassium chloride, potassium nitrate, sodium sulfate, potassium phosphate, ammonium sulfate, ammonium nitrate, sodium carbonate, sodium glutamate, sodium citrate, sodium acetate, and combinations thereof.
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify and/or combine the reference teachings to make the claimed invention by incorporating the methods steps of US20130061351 into the method steps of US20190040407 and incubating the hairy roots in a solution comprising from about 250 mM to about 4 M of a salt as taught by Lallemand et al. where one of ordinary skill in the art before the effective filing date of the claimed invention would have been motivated to do this in order to obtain a method for continuous production and recovery of a secreted compound of interest in optimal amounts by incubating the hairy roots in a solution comprising from about 250 mM to about 4 M of a salt. One of ordinary skill in the art at the time the invention was made would have a reasonable expectation of success because using hairy roots-based expression system for producing a desired compound are known in the art as shown by the above reference teachings. Hence, the claimed invention as a whole is prima facie obvious.
Amending the claims to specifically recite a solution comprising from about 250 mM to about 4 M of a salt selected from a group consisting of sodium salt, potassium salt, chloride salt, sulfate salt, nitrate salt, ammonium salt, phosphate salt, and combinations thereof or, from about 250 mM to about 4 M of a salt selected from the group consisting of sodium chloride, potassium chloride, potassium nitrate, sodium sulfate, potassium phosphate, ammonium sulfate, ammonium nitrate, sodium carbonate, sodium glutamate, sodium citrate, sodium acetate, and combinations thereof would aid in overcoming the rejection.
The reference teachings and rejection have been restated below.
US20190040407 teaches production of protein from hairy roots of a Brassicaceae plant utilizing rhizobium comprising rol genes (see entire publication and claims especially paragraphs [0016]- [0055]. US20190040407 teaches the following in the claims:
1. Method for producing protein of interest from lateral root emergences appearing on hairy roots of a plant belonging to the family of the Brassicaceae, in liquid medium containing at least one auxin, comprising the steps of:
a) transforming a plant belonging to the family of the Brassicaceae with a strain of Rhizobium comprising the rol genes, in order to obtain the hairy roots, and
b) transforming said plant with a vector containing an expression cassette comprising a gene encoding said protein of interest,
the aforesaid steps taking place in a first culture medium, and
c) inducing lateral root emergences on the hairy roots in the presence of at least one auxin in liquid medium constituting a second liquid culture medium, and
d) culture of the hairy roots having lateral root emergences that do not grow longer, and
e) the spontaneous secretion in the medium of the protein of interest during the culture, and
f) the recovery of the aforesaid protein of interest directly from the second liquid culture medium.
2. Method for producing protein of interest from lateral root emergences appearing on hairy roots of a plant belonging to the family of the Brassicaceae, in liquid medium containing at least one auxin, according to claim 1, comprising the steps of:
a) transforming a plant belonging to the family of the Brassicaceae with a strain of Rhizobium comprising the rol genes, in order to obtain the hairy roots, and
b) transforming said plant with a vector containing an expression cassette comprising a gene encoding said protein of interest, and
c) culture in liquid medium of the hairy roots obtained according to steps a) and b), the aforesaid steps taking place in a first culture medium, and
d) inducing lateral root emergences on the hairy roots in the presence of at least one auxin in liquid medium constituting a second liquid culture medium, and
e) culture in the second liquid culture medium, of the hairy roots having lateral root emergences that do not grow longer, and
f) the spontaneous secretion in the second liquid culture medium, of the protein of interest during the culture, and
g) the recovery of the protein of interest directly from the second liquid culture medium.
3. Method for producing protein of interest according to claim 1, in which the strain of Rhizobium is a strain of the species Rhizobium rhizogenes, in particular in which said strain of R. rhizogenes is selected from the strains ATCC 25818, LBA 9402, A4T, A4, LBA1334, ATCC 11325, ATCC 15834 and LMG 155 and is preferentially selected from the strain ATCC 25818 or the strain ICPB TR7.
4. Method for producing protein of interest according to claim 1, in a liquid culture medium containing at least one auxin, from lateral root emergences that do not grow longer appearing on hairy roots of a plant belonging to the family of the Brassicaceae, comprising the steps of:
a) transforming a plant belonging to the family of the Brassicaceae with a strain of Rhizobium comprising the rol genes and an expression cassette comprising a gene encoding said protein of interest in order to obtain the hairy roots, and
b) culture in liquid medium of the hairy roots obtained according to step a),
the aforesaid steps taking place in a first culture medium, and
c) inducing lateral root emergences on the hairy roots in the presence of at least one auxin in liquid medium, constituting a second culture medium, and
d) culture in the second liquid culture medium, of the lateral root emergences that do not grow longer appearing on the aforesaid hairy roots, and
e) the spontaneous secretion in the second liquid culture medium, of the protein of interest during the culture.
5. Method for producing protein of interest according to claim 1, in which the step of inducing lateral root emergences on the hairy roots in the presence of at least one auxin in liquid medium is carried out by the addition of at least one auxin to the first liquid culture medium of the hairy roots, in order to produce the second liquid culture medium.
6. Method for producing protein of interest according to claim 1, in which the step of inducing lateral root emergences on the hairy roots in the presence of at least one auxin in liquid medium is carried out by replacing a first liquid culture medium of the hairy roots with a second liquid culture medium containing at least one auxin.
7. Method for producing protein of interest according to claim 1, in which the step of recovery of the aforesaid expressed protein of interest is carried out directly from the second culture medium after a spontaneous secretion during the culture.
8. Method for producing protein of interest according to claim 1, in which the auxin is selected from: 2,4-dichlorophenoxyacetic acid (2,4-D), 3-indoleacetic acid (IAA), indole-3-butyric acid (IBA), 1-naphthaleneacetic acid (NAA), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), 2,3,5-triiodoacetic acid, 4-chlorophenoxyacetic acid, 2-naphthoxyacetic acid, 1-naphthylacetic acid, 4-amino-3,5,6-trichloropicolinic acid, 3,6-dichloro-2-methoxybenzoic acid (Dicamba) and derivatives thereof by radical modification.
9. Method for producing protein of interest according to claim 1, in which the 2,4-D auxin is used at a concentration from 0.01 to 10 mg/l, in particular from 0.2 to 1 mg/l, in particular 0.5 mg/l, in particular 1 mg/l.
10. Method for producing protein of interest according to claim 1, in which the plant belonging to the family of the Brassicaceae is selected from the genera Arabidopsis, Armoracia, Barbarea, Brassica, Crambe, Eruca, Raphanus, Wasabia and Camelina.
11. Method for producing protein of interest according to claim 1, in which the plant belonging to the family of the Brassicaceae is Brassica rapa or Arabidopsis thaliana.
12. Method for producing protein of interest according to claim 1, in which the protein of interest is a protein of viral origin, in particular a protein of the hepatitis B virus referenced by Swiss-Prot accession number P03141.3, or a protein of animal origin, in particular originating from a mammal, said mammal being selected from the rodents, felines, canines and primates, or a protein of human origin, or a protein of plant origin, in particular the glycosylated plant proteins, lectin or papain.
13. Method for producing protein of interest according to claim 1, in which the protein of interest is selected from the allergens, vaccines, enzymes, enzyme inhibitors, antibodies, antibody fragments, antigens, toxins, antimicrobial peptides, hormones, growth factors, blood proteins, receptors, signalling proteins, protein components of biomedical standards, protein components of cell culture medium, fusion or labelled proteins, cysteine-rich peptides or proteins.
14. Method for producing protein of interest according to claim 1, in which the plant of interest is human gastric lipase.
15. Method for producing lateral root emergences appearing on hairy roots, in liquid medium containing at least one auxin, comprising the steps of:
a) transforming a plant belonging to the family of the Brassicaceae with a strain of Rhizobium comprising the rol genes, in order to obtain the hairy roots, and
b) inducing lateral root emergences that do not grow longer on the hairy roots in the presence of at least one auxin in liquid medium.
16. The method of claim 1, further comprising a step of recovery of the aforesaid protein of interest that has accumulated in the tissues by grinding the hairy roots.
17. The method of claim 2, further comprising a step of recovery of the aforesaid protein of interest that has accumulated in the tissues by grinding the hairy roots.
18. The method of claim 4, further comprising a step of recovery of the aforesaid expressed protein of interest by extraction from the tissues and directly from the second liquid culture medium.
19. The method of claim 7, wherein the step of recovery of the aforesaid expressed protein of interest is carried out by grinding of the cultured tissues.
20. Method for producing protein of interest according to claim 2, in which the strain of Rhizobium is a strain of the species Rhizobium rhizogenes, in particular in which said strain of R. rhizogenes is selected from the strains ATCC 25818, LBA 9402, A4T, A4, LBA1334, ATCC 11325, ATCC 15834 and LMG 155 and is preferentially selected from the strain ATCC 25818 or the strain ICPB TR7.
The teachings of the reference differ from the claims in that the reference does not teach incubating the hairy roots in a solution comprising from about 250 mM to about 4 M of a salt.
US20130061351 teaches methods for producing recombinant proteins from plant hairy roots (see entire publication and claims especially paragraphs [0007]-[0020]. US20130061351 teaches the following in the claims:
16. A method for obtaining a recombinant protein from hairy roots comprising the steps of:
a) transforming a plant with a strain of Agrobacterium rhizogenes and/or with a strain of Agrobacterium tumefaciens comprising the rol genes;
and
b) transforming said plant with a vector containing an expression cassette comprising a gene encoding said recombinant protein;
wherein said plant belongs to the Brassicaceae family.
17. A method according to claim 16, wherein said expression cassette comprises a signal peptide.
18. A method according to claim 16, wherein step a) and step b) are performed simultaneously by transforming said plant belonging to the Brassicaceae family with a strain of Agrobacterium rhizogenes, wherein said strain of Agrobacterium rhizogenes contains an expression cassette comprising a gene encoding said recombinant protein.
19. A method according to claim 16, wherein said plant belonging to the Brassicaceae family is selected from the group consisting of Raphanus sativus, Raphanus sativus var. niger, Brassica oleracea L. convar and Brassica rapa.
20. A method according to claim 16, wherein said plant belonging to the Brassicaceae family is Brassica rapa.
21. A method according to claim 16, wherein said strain of Agrobacterium rhizogenes is the strain ATCC 25818.
22. A method according to claim 16, wherein said expression cassette comprises a promoter, a signal peptide, a gene encoding said recombinant protein and a polyadenylation sequence.
23. A method according to claim 22, wherein said promoter is a promoter derived from a virus infecting Brassicaceac plants.
24. A method according to claim 22, wherein said promoter is the cauliflower mosaic virus 35S (CaMV35S) promoter.
25. A method according to claim 22, wherein said signal peptide is derived from a Brassicaceae plant.
26. A method according to claim 22, wherein said signal peptide is a signal peptide from the Arabidopsis pectin methylesterase AT1G69940 signal peptide or a variant thereof, such as the signal peptide as set forth in SEQ ID NO:1.
27. A method according to claim 16, wherein said recombinant protein is selected from the group consisting of allergens, vaccines, enzymes, enzyme inhibitors, antibodies, antibody fragments, antigens, toxins, anti-microbial peptides, hormones, growth factors, blood proteins, receptors and signaling proteins, protein component of biomedical standards, protein component of cell culture media, fusion or tagged proteins, cystein (disulfide bridges)-rich peptides and proteins, and plant proteins.
28. A method according to claim 27, wherein said blood proteins are albumin, coagulation factors, or transferrin.
29. A method according to claim 27, wherein said plant proteins are lectins or papain.
30. A hairy root culture obtainable by transforming a plant with a strain of Agrobacterium rhizogenes, wherein said plant belongs to the Brassicaceae family and wherein said strain of Agrobacterium rhizogenes contains an expression cassette comprising a gene encoding said recombinant protein.
31. A culture medium containing a recombinant protein obtainable by the method according to claim 16.
32. A recombinant protein obtainable by the method according to claim 16.
Lallemand et al. teach extracellular peptidase hunting for improvement of protein production in plant cells and roots, and use of NaCl at 1 M under agitation for 1 h at 4°C to recover extracellular compounds produced in plant roots (p. 2 : Rhizosecretome harvesting). See entire publication and abstract especially MATERIALS AND METHODS section, and pages 2-3.
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify and/or combine the reference teachings to make the claimed invention by incorporating the methods steps of US20130061351 into the method steps of US20190040407 and incubating the hairy roots in a solution comprising from about 250 mM to about 4 M of a salt as taught by Lallemand et al. One of ordinary skill in the art before the effective filing date of the claimed invention would have been motivated to do this in order to obtain a method for recovering a secreted compound of interest in optimal amounts by incubating the hairy roots in a solution comprising from about 250 mM to about 4 M of a salt. It would have been obvious to adjust the method steps conditions, incubation time and temperature, and amounts of salts recited in the claims as routine optimization and/or as desired for optimal production of the desired secreted compound. One of ordinary skill in the art at the time the invention was made would have a reasonable expectation of success because using hairy roots-based expression system for producing a desired compound are known in the art as shown by the above reference teachings. Hence, the claimed invention as a whole is prima facie obvious.
Conclusion
No claim is allowed.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any extension fee pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Christian L Fronda whose telephone number is (571)272 0929. The examiner can normally be reached Monday-Thursday and alternate Fridays between 9:00AM-5:00PM.
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/CHRISTIAN L FRONDA/Primary Examiner, Art Unit 1652