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 .
The Amendments and Remarks filed 6/1/26 in response to the Office Action of 3/4/26 are acknowledged and have been entered.
Claims 63-67 have been added by Applicant.
Claims 29-31, 33-41, 45, 61, and 63-67 are pending.
Claims 29, 33, 39, 40, 45, and 61 have been amended by Applicant.
Claims 29-31, 33-41, 45, 61, and 63-67 are currently under examination.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
The following Office Action contains NEW GROUNDS of rejections Necessitated by Amendments.
Rejections Withdrawn
All previous rejections are withdrawn.
New Rejections Necessitated by Amendments
Claim Rejections - 35 USC § 112
Claims 30, 33, 45, 61, and 63-67 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 applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 30 is rejected for reciting “The method of claim 29, wherein the selected cell line is then cultured at the temperature regime that resulted in reduced expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain.” Claim 29 recites culturing at least two cell lines under either a first temperature regime or a second temperature regime and then “c) comparing the expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain produced by cell cultured at each temperature regime; and d) selecting a cell line that resulted in reduced expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain at each temperature regime.” Therefore, claim 29 encompasses methods where the selected cell line exhibited reduced unpaired impurity at a first temperature regime (as compared to the second temperature regime) and reduced mis-paired long heavy chain at the second temperature regime (are compared to the first temperature regime). However, the metes-and-bounds of claim 30 are unclear because it is unclear which temperature regime of claim 29 is “the temperature regime” that resulted in reduced expression of at least at least one product-related impurity comprising an unpaired or mis-paired long heavy chain – because claim 29 recites the selected cell line reduced expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain in both temperature regimes (see “at each temperature regime” at the last lines of claim 29).
Claim 33 is rejected for reciting “The method according to claim 1, wherein….” Claim 1 has been cancelled. The metes-and-bounds of claim 33 are unclear because it is unclear which method claim 33 is further limiting. Further, there is a lack of antecedent basis for “The method according to claim1” in claim 33.
Claim 45 is rejected for reciting a method of culturing at least one cell culture from each or two or more clonally derived cultures that both recombinantly express an asymmetric multispecific antibody at a first temperature regime (single temperature) and a second temperature regime (temperature shift), and then “d) comparing the amount of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain produced by the cells cultured at each temperature regime; e) selecting a clonally derived culture that modulated production of the product related impurity at each temperature regime; f) establishing a cell line expressing the asymmetric multispecific antibody from the selected clonally derived culture; g) inoculating a bioreactor with the cell line expressing the asymmetric multispecific antibody; h) culturing the cells to express the asymmetric multispecific antibody at the temperature regime that modulated the production of the product-related impurity…..” The metes-and-bounds of claim 45 are unclear because (i) it is unclear which product related impurity is “the product related impurity” at each temperature regime at step “e)” and (ii) it is unclear which temperature regime is “the temperature regime” that modulated the production of the product-related impurity at step “h” – because step e) of claim 45 recites the selected cell line modulated production of the product related impurity in both temperature regimes (see “at each temperature regime” at step e) of claim 45).
Claims 61 and 63-67 are rejected because claim 1 recites a method comprising culturing cell lines in a temperature regime comprising a first temperature and then shifting the cells to a lower temperature and then “(e) selecting a cell line that resulted in reduced expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain at each temperature regime….” The metes-and-bounds of claims are unclear because the claims only recite a single (temperature shift) temperature regime – therefore, it is unclear what is meant by “each temperature regime.”
Claim Rejections - 35 USC § 103
Claim(s) 29-31, 33-41, and 45 is/are rejected under 35 U.S.C. 103 as being unpatentable over Gomez et al (Biotechnology and Bioengineering, 2018, 115(12): 2930-2940; 9/28/23 IDS) in view of Mchugh et al (Biotechnology Progress, 2020, 36(e2959): 1-11), Torres et al (PLOSOne, 2018, 13(3)(e0194510): 1-23), Ali et al (WO 2019/140196 A1; 7/18/2019; 9/28/23 IDS).
Gomez et al teaches a biphasic cell culture process for generating asymmetric multispecific antibodies comprising a “temperature shift” cell culture temperature regime that comprises culturing stably transformed clonally derived CHO host cells (“cell line X”) expressing the asymmetric multispecific antibody in a vessel comprising a standard culture medium at 36 °C to maximize cell growth first for 3 days and then shifting to culturing the cells at 32.5 °C, and harvesting of the asymmetric multispecific antibody product on days 3, 6, 8, 10, and 13 for analysis, followed by protein-A or CEX chromatography purification of the asymmetric multispecific antibody (see 2.1 Cell culture at right column on page 2931, in particular). Gomez et al further teaches the biphasic cell culture process for generating asymmetric multispecific antibodies comprising a “temperature shift” cell culture temperature regime with the clonally derived CHO host cells (“cell line X”) expressing the asymmetric multispecific antibody as an “optimal process” that led to higher yield and “improve quality” (see Table 4, and left column on page 2939, in particular). Table 4 of Gomez et al further illustrate that the clonally derived CHO host cells (“cell line X”) expressing the asymmetric multispecific antibody cultured with the “temperature shift” temperature regimen and purified with protein-A resulted in higher yields and strikingly reduced expression of both “Half B-Ab” impurity (comprising an unpaired heavy chain) and LMW impurity as compared to clonally derived CHO host cells (“cell line X”) expressing the asymmetric multispecific antibody cultured at a constant temperature regimen and purified with protein-A (see Table 4 and Figure 4 for description of “Half B-Ab”, in particular).
While the clonally derived CHO host cells (“cell line X”) expressing the asymmetric multispecific antibody cultured with the “temperature shift” temperature regimen purified with protein-A or CEX resulted in similar “HMW” impurities (comprising a mis-paired long heavy chain) as compared to clonally derived CHO host cells (“cell line X”) expressing the asymmetric multispecific antibody cultured at a constant temperature regimen and purified with protein-A or CEX, Gomez does teach a slightly higher level of the HMW impurity with the constant temperature regimen than the temperature shift regimen purified with protein-A as compared to the cells cultured at a constant temperature regime and purified by protein-A (see Table 4 and pages 2936-2937 for description of “HMW”, in particular). See Figure 4:
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Gomez et al further teaches effects of temperature regimes on other clonally derived CHO host cells expressing the same asymmetric multispecific antibody as “cell line X” did not mirror those exhibited by “cell line X” (page 2938, in particular).
Gomez et al does not explicitly teach recited “selecting” steps or generating an asymmetric multispecific antibody comprising an IL-21 mutein. However, these deficiencies are made up in the teachings of Mchugh et al , Torres et al, and Ali et al.
Mchugh et al teaches temperature shifts to lower culture temperatures are frequently employed in manufacturing of protein therapeutics in mammalian cells to improve productivity, viability, and quality attributes that the extent to which a temperature shift affects productivity and quality may vary depending on the expression host and characteristics of the expressed protein (Abstract, in particular). Mchugh et al further teaches two CHO clones expressing different monoclonal antibodies respond differently to a temperature shift despite having a parental CHO cell line (Abstract, in particular).
Tores et al teaches two clonally derived CHO cell lines (CN1 and CN2) that recombinantly express the same antibody (anti-TNFa) differ from one another, in viability and antibody production, when clonally derived CHO cell lines are cultured at different temperatures (Fig. 1, in particular). A lower temperature seemed to increase antibody production and anti-TNFa mRNA expression to a greater extent in the CN2 clone, as compared to the CN1 clone (Fig. 1 and Fig. 3, in particular).
Ali et al teaches an asymmetric multispecific antibody comprising a PD-1 targeting antibody fused to an IL-21 mutein that overcomes significant barriers associated with cytokine therapeutics and allows for antibody-like dosing and selectively delivery of the IL-21 mutein in a PD-1 targeted manner wherein the IL-21 mutein can selectively activate and expand PD-1 expressing T cells in vivo ([0089], in particular). The asymmetric multispecific antibody of Ali et al has the following general structure:
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Ali et al further teaches the multispecific antibody comprising VH and VL domains of SEQ ID NOs: 382-384 and 385-387, which comprise instant SEQ ID NOs: 382-384 and 385-389 ([00227], in particular). Ali et al further teaches the multispecific antibody comprising VH and VL domains of SEQ ID NOs: 362-364 and 365-369, which comprise instant SEQ ID NOs: 362-364 and 365-369 ([00237], in particular). Ali et al further teaches the multispecific antibody comprising VL domain of SEQ ID NOs: SEQ ID NO:391 (which comprise instant SEQ ID NO: 391), a heavy chain comprising SEQ ID NO: SEQ ID NO:556 (which comprises instant SEQ ID NO:556), and a heavy chain attached to IL-21 mutein comprising SEQ ID NO:501 (which comprises instant SEQ ID NO:501) ([00227], in particular). Ali et al further teaches the multispecific antibody comprising VL domain of SEQ ID NO: 371 (which comprise instant SEQ ID NO: 371), a heavy chain comprising SEQ ID NO: 559 (which comprises instant SEQ ID NO:559), and a heavy chain attached to IL-21 mutein comprising SEQ ID NO:513 (which comprises instant SEQ ID NO:513 and instant SEQ ID NO:244) ([00237], in particular).
One of ordinary skill in the art would have been motivated, with a reasonable expectation of success, to perform a combined method of identifying a clonal cell line that can optimally generate the asymmetric multispecific antibody of Ali et al under optimized culture conditions comprising (i) establishing at least two clonally derived cell lines expressing the asymmetric multispecific antibody Ali et al, (ii) culturing at least one cell culture of each clonally derived cell lines at a first temperature (temperature-shift) regime at 36 °C for three days and then shifting the culture temperature to 32.5 °C and analyzing the antibody product on days 3, 6, 8, 10, and 13 (as taught by Gomez et al), (iii) culturing one other cell culture of each clonally derived cell lines continuously at a temperature regime of 36 °C (continuous temperature regime of Gomez et al), (iv) purifying the expression products by chromatography and comparing the expression of product-related asymmetric antibody production impurities of Gomez et al (including unpaired or mis-paired long heavy chains) for each clonally derived cell lines at each temperature regimen, and (v) selecting a clonally derived cell line that exhibits a culture condition that results in optimal asymmetric multispecific antibody production with reduced expression of product-related impurities of unpaired or mis-paired long heavy chains (as taught by Gomez et al) and then culturing the selected clonally derived cell line in the culture condition that results in optimal asymmetric multispecific antibody production for the selected clonally derived cell line in an effort to optimize generation of the asymmetric multispecific antibody of Ali et al. The combined method evaluates multiple clonally derived cell lines with each temperature regimen because Mchugh et al teaches temperature shifts to lower culture temperatures are frequently employed in manufacturing of protein therapeutics in mammalian cells to improve productivity, viability, and quality attributes that the extent to which a temperature shift affects productivity and quality may vary depending on the expression host and characteristics of the expressed protein (Abstract, in particular), Tores et al teaches two clonally derived CHO cell lines (CN1 and CN2) that recombinantly express the same antibody (anti-TNFa) differ from one another (sometimes slightly), in viability and antibody production, when clonally derived CHO cell lines are cultured at different temperatures (Fig. 1, in particular), and Gomez et al teaches effects of temperature regimes on other clonally derived CHO host cells expressing the same asymmetric multispecific antibody as “cell line X” did not mirror those exhibited by “cell line X” (page 2938, in particular).
Further, it is predictable that selected clonally derived cell lines of embodiments of the combined method include those that exhibit a culture condition that results in optimal asymmetric multispecific antibody production that may exhibit strikingly reduced expression of one product-related impurities of unpaired or mis-paired long heavy chains under said culture condition (such as temperature shift regime), but may have a bit higher level of a different product-related impurity at other conditions (such as continuous temperature regime), as exhibited by “cell line X” of Gomez et al.
Further, one of ordinary skill in the art would have been motivated, with a reasonable expectation of success, to perform said combined method wherein the selected clonally derived cell line that exhibits a culture condition that results in optimal asymmetric multispecific antibody production with reduced expression of product-related impurities of unpaired or mis-paired long heavy chains is inoculated into a bioreactor (such as a bioreactor comprising a shake flask of page 2931 of Gomez et al), culture the selected clonally derived cell line under said culture condition that results in optimal asymmetric multispecific antibody production, and harvest and chomotographically purify the multispecific antibody (as taught by Gomez et al) in order to obtain a purified the PD-1 targeting antibody of Ali et al fused to an IL-21 mutein that overcomes significant barriers associated with cytokine therapeutics and allows for antibody-like dosing and selectively delivery of the IL-21 mutein in a PD-1 targeted manner wherein the IL-21 mutein can selectively activate and expand PD-1 expressing T cells in vivo ([0089] of Ali et al, in particular)
This is an example of use of known technique (optimization by temperature-shift) to improve similar methods or products in the same way. This is further an example of some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. Therefore, the invention as a whole would have been prima facie obvious to one of ordinary skill in the art, absent unexpected results.
Claim Rejections - 35 USC § 103
Claim(s) 29-31, 33-41, 45, 61, and 63-67 is/are rejected under 35 U.S.C. 103 as being unpatentable over Gomez et al (Biotechnology and Bioengineering, 2018, 115(12): 2930-2940; 9/28/23 IDS) in view of Mchugh et al (Biotechnology Progress, 2020, 36(e2959): 1-11), Torres et al (PLOSOne, 2018, 13(3)(e0194510): 1-23), Ali et al (WO 2019/140196 A1; 7/18/2019; 9/28/23 IDS), as applied to claims 29-31, 33-41, and 45 above, and further in view of Lionberger et al (WO 2019/075476 A2; 4/18/19).
Teachings of Gomez et al, Mchugh et al, Torres et al, and Ali et al are discussed above.
Gomez et al, Mchugh et al, Torres et al, and Ali et al do not specifically teach methods involving a nanofluidic chip. However, these deficiencies are made up in the teachings of Linoberger et al.
Lionberger et al teaches developing a new antibody production line can take many months of work and cost millions of dollars ([00219], in particular). Lionberger et al further teaches a microfluidic device (same as “chip”; see [00060]) that is able to screen and identify promising clones days after seeding individual clones that would offers significant time and cost advantages ([00219], in particular). Lionberger et further teaches the microfluidic devices comprising many (including 3,500) chambers ([00059] and [00510], in particular) in which analytes can be produced and measured ([0002], in particular). Lionberger et al further teaches analytes that are antibodies can be measured bound to a reporter that is a peptide epitope that binds the antibody ([00251], in particular). Lionberger et further teaches top analyte producers can be selected and exported from the microfluidic devices in order to expand the cells (“micro-objects”) ([0008], in particular). Lionberger et further teaches cells expressing analytes can be exported into well plates those wells can then be introduced to shaker flasks and then scaled-up ([00500], in particular). Lionberger et further teaches the temperature of the microfluidic device can be regulated ([00217], in particular).
One of ordinary skill in the art would have been motivated, with a reasonable expectation of success, to rapidly develop an optimized clone that generates an asymmetric multispecific antibody of Ali et al by performing the combined method of Gomez et al, Mchugh et al, Torres et al, and Ali et al at wherein many different cultured clonally-derived cell lines expressing the antibody of Ali et al are plated as single cell clones cultured in many distinct chambers of a microfluidic device of Lioberger et al, wherein the cells are cultured in medium at the temperatures and time points of the combined method, and amounts of asymmetric multispecific antibody products produced by each clone are determined at time points of the combined method, and those clonally derived cells that exhibits a culture condition that results in optimal asymmetric multispecific antibody production with reduced expression of product-related impurities of unpaired or mis-paired long heavy chains (as taught by Gomez et al) are selected and exported (as taught by Lionberger et al) any time cells that are identified as exhibiting a culture condition that results in optimal asymmetric multispecific antibody production – including after day 6 (after the three days at lower temperature of the combined method) into multi-wells of plates (such as those described on right column on page 2931 of Gomez et al), shaker flasks, and then scaled-up because the microfluidic device of Lionberger et al has the ability to culture antibody-producing cells at optimized temperatures and has the benefit of analyzing thousands of clones at a time for optimal antibody production. This is an example of prior art elements according to known methods to yield predictable results. This is further an example of use of known technique to improve similar methods in the same way. This is further drawn to some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to combine prior art reference teachings to arrive at the claimed invention.
Therefore, the invention as a whole would have been prima facie obvious to one of ordinary skill in the art, absent unexpected results.
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claim 29-31, 33-41, 45, 61, and 63-67 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention. This is a NEW MATTER rejection.
Claims 29-31 and 34-41 are rejected because step d) of claim 29 recites a method comprising culturing at least two cell lines, wherein a culture from each cell line is cultured at first temperature regime and second (temperature shift) temperature regime and selecting a cell line that resulted in reduced expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain at each temperature regime. Descriptions of methods comprising culturing at least two cell lines, wherein a culture from each cell line is cultured at first temperature regime and second (temperature shift) temperature regime and selecting a cell line that resulted in reduced expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain at each temperature regime are not found in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventors, at the time the invention was filed, had possession of the claimed invention.
Claim 45 recites a method of culturing at least one cell culture from each or two or more clonally derived cultures that both recombinantly express an asymmetric multispecific antibody at a first temperature regime (single temperature) and a second temperature regime (temperature shift), and then “d) comparing the amount of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain produced by the cells cultured at each temperature regime; e) selecting a clonally derived culture that modulated production of the product related impurity at each temperature regime; f) establishing a cell line expressing the asymmetric multispecific antibody from the selected clonally derived culture; g) inoculating a bioreactor with the cell line expressing the asymmetric multispecific antibody; h) culturing the cells to express the asymmetric multispecific antibody at the temperature regime that modulated the production of the product-related impurity…..”. Descriptions of methods of culturing at least one cell culture from each or two or more clonally derived cultures that both recombinantly express an asymmetric multispecific antibody at a first temperature regime (single temperature) and a second temperature regime (temperature shift), and then “d) comparing the amount of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain produced by the cells cultured at each temperature regime; e) selecting a clonally derived culture that modulated production of the product related impurity at each temperature regime; f) establishing a cell line expressing the asymmetric multispecific antibody from the selected clonally derived culture; g) inoculating a bioreactor with the cell line expressing the asymmetric multispecific antibody; h) culturing the cells to express the asymmetric multispecific antibody at the temperature regime that modulated the production of the product-related impurity…..” are not found in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventors, at the time the invention was filed, had possession of the claimed invention.
Claims 61 and 63-67 are rejected because claim 1 recites a method comprising culturing cell lines in a temperature regime comprising a first temperature and then shifting the cells to a lower temperature and then “(e) selecting a cell line that resulted in reduced expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain at each temperature regime….” Descriptions of methods comprising culturing cell lines in a temperature regime comprising a first temperature and then shifting the cells to a lower temperature and then “(e) selecting a cell line that resulted in reduced expression of at least one product-related impurity comprising an unpaired or mis-paired long heavy chain at each temperature regime….” are not found in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventors, at the time the invention was filed, had possession of the claimed invention.
Conclusion
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 nonprovisional extension fee (37 CFR 1.17(a)) 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 mailing date of this final action.
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/SEAN E AEDER/ Primary Examiner, Art Unit 1642
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