|Table of Contents|

Bispecific Anti-CD3×Anti-CD87 Single-Chain Antibody Mediates CytolyticActivity of Mononuclear Cells Against CD87-Positive Prostate Cancer in vitro(PDF)

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2017年02期
Page:
51-
Research Field:
·生命科学·
Publishing date:

Info

Title:
Bispecific Anti-CD3×Anti-CD87 Single-Chain Antibody Mediates CytolyticActivity of Mononuclear Cells Against CD87-Positive Prostate Cancer in vitro
Author(s):
Gao Wanli; Yang Li; Liu Shan; Zhao Zhihui
School of Life Sciences,Nanjing Normal University,Institute of Biochemistry and Biological Products,Jiangsu Key Laboratory for Molecular and Medical Biotechnology,Nanjing 210023,China
Keywords:
single chain bispecific antibody; cytotoxity; prostate cancer; prokaryotic expression
PACS:
Q819
DOI:
10.3969/j.issn.1001-4616.2017.02.009
Abstract:
In this study,the aim was to investigate whether anti-CD3×anti-CD87 single-chain bi-functional specific antibody(scBsAb)could mediate the cytotoxicity of peripheral blood mononuclear cells(PBMCs)against CD87-positive prostatic cancer cells. First,the prokaryotic expression vector of pET24a/scBsAb was constructed and transformed into competent cells of E.coil BL-21(DE3)by genetic engineering technique. After induction with IPTG,the bacteria were ultrasonicated,and scBsAb in the form of inclusion bodies were harvested by centrifugation. The scBsAb was further purified by affinity chromatography under denaturing conditions and refolded by dilution. Second,the antigens binding activity of scBsAb was examined by flow cytometry. Lastly,the scBsAb-mediated cytotoxicity of PBMCs against CD87-positive prostatic cancer cell line PC-3 was determined by using CCK-8 assay. Our results showed that the scBsAb with high purity was obtained. scBsAb could both specifically bind to CD87 and CD3. The scBsAb-mediated killing of PBMCs against CD87+ PC-3 cells was related to the concentration of used scBsAb and the ratio of effector to target(E:T). When E:T was 10:1 and the scBsAb concentration was 100 μg/mL,the killing rate was 40.86%; whereas when the E:T ratio was 40:1 and scBsAb concentration was 100 μg/mL,the killing rate was 60.9%. In addition,ELISA assay showed that IFN-γ level in the co-culture system was significantly increased during the killing process. Our data indicate that anti-CD3×anti-CD87 scBsAb could mediate PBMCs to kill CD87-positive tumor cells in vitro.

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Last Update: 2017-06-30