Preparation and identification of human soluble sPD-L1 and its antibodies.
- Author:
Li-Hui XU
1
;
Xiao-Yun CHI
;
Feng-Yao LI
;
Qian-Tao JIA
;
Qing-Bing ZHA
;
Xian-Hui HE
Author Information
1. Institute of Tissue Transplantation and Immunology, College of Life Science and Technology, Jinan University, Guangzhou 510632, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies;
immunology;
isolation & purification;
Antigens, CD;
genetics;
immunology;
metabolism;
B7-H1 Antigen;
Chromatography, Affinity;
Electrophoresis, Polyacrylamide Gel;
Enzyme-Linked Immunosorbent Assay;
Escherichia coli;
genetics;
Female;
Humans;
Immune Sera;
immunology;
Immunoblotting;
Mice;
Mice, Inbred C57BL;
Recombinant Proteins;
immunology;
isolation & purification;
metabolism;
Solubility
- From:
Chinese Journal of Biotechnology
2007;23(1):106-111
- CountryChina
- Language:Chinese
-
Abstract:
This study reports the preparation and identification of soluble programmed death-1 (PD-1) ligand-1 (sPD-L1) and its antibodies of mouse origin. Immobilized metal ion affinity chromatography was used to perform on-column refolding with simultaneous purification of denatured sPD-L1, and soluble sPD-L1 with purity of 95% was obtained. The purified sPD-L1 was verified by immunoblotting using a commercial goat-anti-human PD-L1 antibody. An ELISA-based assay showed that it also had high binding activity for its cognate receptor PD-1. Furthermore, mouse anti-sPD-L1 antiserum of high titer was raised using the purified sPD-L1 as an immunogen, and the specific IgG antibodies were purified using sPD-L1-HiTrap affinity chromatography. In addition, a sensitive sandwich ELISA was established using the purified IgG antibodies together with the commercial goat antibodies. In conclusion, the preparation of soluble sPD-Ll and its antibodies provide the basis for detection of the potential anti-PD-L1 antibodies and soluble PD-L1 in humans as well as for further investigation of its in vivo bioactivities and characterization of its potential receptors.
