Construction, expression, and identification of the gene of human anti-prostate specific membrane antigen single-chain antibody / 中华男科学杂志
National Journal of Andrology
; (12): 1063-1067, 2014.
Article
in Zh
| WPRIM
| ID: wpr-319567
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct, express and purify human fusion proteins composed of a single-chain antibody fragment scFv that recognizes the prostate specific membrane antigen (PSMA) protein, Fdt, HA2 and tp, and to analyze the binding activity of the expressed fusion proteins.</p><p><b>METHODS</b>The fusion protein genes scFv, scFv-tp, and scFv-Fdt-HA2-tp were amplified by PCR, and the genes obtained were then cloned into the expression vector pET28 and expressed in E. coli BL21. The expressed products were identified by SDS-PAGE and Western blot and purified with Ni(2+)-NTA chelating agarose. The antigen-binding activity of the fusion proteins was determined by ELISA.</p><p><b>RESULTS</b>The human anti-PSMA fusion gene was successfully constructed and expressed in M15 as the inclusion body after induced with IPTG. All the target proteins expressed could bind the PSMA antigen.</p><p><b>CONCLUSION</b>Fusion proteins can specifically bind the PSMA antigen. This finding contributes to the study of the targeted delivery of siRNA.</p>
Full text:
1
Database:
WPRIM
Main subject:
Recombinant Fusion Proteins
/
Polymerase Chain Reaction
/
Cloning, Molecular
/
RNA, Small Interfering
/
Glutamate Carboxypeptidase II
/
Electrophoresis, Polyacrylamide Gel
/
Allergy and Immunology
/
Escherichia coli
/
Single-Chain Antibodies
/
Genetics
Type of study:
Prognostic_studies
Limits:
Humans
/
Male
Language:
Zh
Journal:
National Journal of Andrology
Year:
2014
Document type:
Article