Simultaneous expression of displayed and secreted antibodies for antibody screen.
The display of full-length antibody on the cell surface was achieved by fusing a transmembrane domain of the platelet-derived growth factor receptor (PDGFR) to the C-terminus of the heavy chain constant region. We also incorporated a furin cleavage site between the constant region and PDGFR transmembrane domain to obtain secreted antibodies. As a result, antibodies can be expressed simultaneously on the cell surface in a membrane-anchored version for screening and selecting through fluorescence-activated cell sorting (FACS) analysis, as well as in conditioned medium in a secreted version for function analysis.
Nanfang Hospital, Southern Medical University, Guangzhou, China.
Yuanping
Y
Zhou
Junjie
J
Wang
Ivan
I
Zhou
Haibo
H
Lou
Chang-Zheng
CZ
Li
Zhen-Rui
ZR
Chen
Zhe-Huan
ZH
Zhang
Shuwen
S
Liu
Shuguang
S
Wu
Wanlong
W
Tan
Shibo
S
Jiang
Chen
C
Zhou
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D003412:Cricetulus; D004262:DNA Restriction Enzymes / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D045683:Furin / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry*; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015202:Protein Engineering; D059748:Proteolysis; D017479:Receptors, Platelet-Derived Growth Factor / Q000235:genetics* / Q000378:metabolism
D000906:Antibodies; D007143:Immunoglobulin Heavy Chains; D017479:Receptors, Platelet-Derived Growth Factor; D004262:DNA Restriction Enzymes; C468731:FURIN protein, human; D045683:Furin