US 12,241,178 B2
Recombinant fusion proteins and libraries from immune cell repertoires
David Scott Johnson, San Francisco, CA (US); Adam Adler, Belmont, CA (US); and Rena Mizrahi, Pacifica, CA (US)
Assigned to GigaGen, Inc., San Carlos, CA (US)
Filed by GigaGen, Inc., South San Francisco, CA (US)
Filed on Jul. 11, 2022, as Appl. No. 17/862,101.
Application 17/862,101 is a division of application No. 16/886,612, filed on May 28, 2020, granted, now 11,702,765.
Application 15/156,214 is a division of application No. 14/734,953, filed on Jun. 9, 2015, granted, now 9,422,547, issued on Aug. 23, 2016.
Application 16/886,612 is a continuation of application No. 16/399,759, filed on Apr. 30, 2019, granted, now 10,689,641, issued on Jun. 23, 2020.
Application 16/399,759 is a continuation of application No. 15/156,214, filed on May 16, 2016, abandoned.
Prior Publication US 2023/0070140 A1, Mar. 9, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C40B 40/08 (2006.01); C07K 14/725 (2006.01); C07K 16/00 (2006.01); C12N 15/10 (2006.01); C12Q 1/6876 (2018.01); C40B 50/06 (2006.01)
CPC C40B 40/08 (2013.01) [C07K 14/7051 (2013.01); C07K 16/00 (2013.01); C12N 15/1037 (2013.01); C12N 15/1075 (2013.01); C12N 15/1093 (2013.01); C12Q 1/6876 (2013.01); C07K 2317/14 (2013.01); C07K 2317/622 (2013.01); C40B 50/06 (2013.01)] 12 Claims
OG exemplary drawing
 
1. A plurality of expression vectors comprising at least 1,000 unique recombinant fusion polynucleotides, wherein each recombinant fusion polynucleotide encodes a full-length recombinant immunoglobulin and comprises:
a. a first polynucleotide and a second polynucleotide and a linker polynucleotide,
each first polynucleotide encoding a heavy chain variable domain from a cognate pair from a single isolated mammalian B cell and an IgG heavy chain constant domain fragment; and
each second polynucleotide encoding a light chain variable domain from the cognate pair from the single isolated mammalian B cell and an IgK light chain constant domain; and
the linker polynucleotide linking the first and second polynucleotides;
b. a third recombinant polynucleotide linked to the recombinant fusion polynucleotides, comprising a first promoter sequence and a sequence encoding a portion of the IgG heavy chain constant domain; and
c. a fourth recombinant polynucleotide linked to the recombinant fusion polynucleotides or the third recombinant polynucleotide, comprising a second promoter sequence or a translational skip site;
wherein the polynucleotides of a. b. and c. are linked via Gibson assembly; and
the expression vectors are plasmids or phagemids.