Preparation and identification of human soluble sPD-L1 and its antibodies / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 106-111, 2007.
Article
in Chinese
| WPRIM
| ID: wpr-325410
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Solubility
/
Recombinant Proteins
/
Enzyme-Linked Immunosorbent Assay
/
Immunoblotting
/
Antigens, CD
/
Chromatography, Affinity
/
Electrophoresis, Polyacrylamide Gel
/
Allergy and Immunology
/
Escherichia coli
/
B7-H1 Antigen
Type of study:
Diagnostic study
Limits:
Animals
/
Female
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2007
Type:
Article
Similar
MEDLINE
...
LILACS
LIS