	US 12,404,512 B2
	Method for optimizing antibody expression
Md Harunur Rashid, La Jolla, CA (US); Mark Shimazu, La Jolla, CA (US); and Feng Tian, La Jolla, CA (US)
Assigned to Ambrx, Inc., La Jolla, CA (US)
Appl. No. 17/051,690
Filed by Ambrx, Inc., La Jolla, CA (US)
PCT Filed May 1, 2019, PCT No. PCT/US2019/030295 § 371(c)(1), (2) Date Oct. 29, 2020, PCT Pub. No. WO2019/213331, PCT Pub. Date Nov. 7, 2019.
Claims priority of provisional application 62/665,093, filed on May 1, 2018.
Prior Publication US 2021/0108213 A1, Apr. 15, 2021
Int. Cl. C12N 15/63 (2006.01); C07K 14/245 (2006.01); C07K 14/535 (2006.01); C07K 16/28 (2006.01); C12N 9/90 (2006.01); C12N 15/67 (2006.01); C12N 15/70 (2006.01)

CPC C12N 15/67 (2013.01) [C07K 14/245 (2013.01); C07K 16/2809 (2013.01); C12N 9/90 (2013.01); C07K 2317/522 (2013.01); C07K 2317/55 (2013.01); C07K 2317/92 (2013.01); C12Y 502/01008 (2013.01)]

22 Claims

1. A vector for expressing a recombinant protein comprising a site-specifically incorporated non-naturally encoded amino acid in Escherichia coli (E. coli), comprising: (i) a nucleic acid sequence encoding the recombinant protein; and (ii) a nucleic acid sequence comprising a partition B (parB) locus having the nucleobase sequence of SEQ ID NO: 12, wherein the recombinant protein is an antibody or an antibody fragment, and wherein the parB locus is positioned in:

a) a reverse transcriptional orientation and upstream relative to the nucleic acid sequence encoding the recombinant protein;

b) a forward transcriptional orientation and downstream relative to the nucleic acid sequence encoding the recombinant protein; or

c) a reverse transcriptional orientation and downstream relative to the nucleic acid sequence encoding the recombinant protein.