Study on the construction and expression of anti-CD3/anti-CD20 Diabody
- VernacularTitle:微型双功能抗体抗CD3/抗CD20的构建和表达
- Author:
Dongsheng XIONG
;
Yuanfu XU
;
Chunzheng YANG
;
Al ET
- Publication Type:Journal Article
- Keywords:
Monoclonal antibody CD3 CD20 Engineering antibody Bispecific antibody
- From:
Chinese Journal of Immunology
2001;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct and express anti CD3/anti CD20 Diabody and identify its biological activity.Methods:PCR and overlap PCR were used to construct anti CD3/anti CD20 Diabody.DNA sequence was analyzed by the Terminus of Dideoxy Nucleotide.The product was purified by affinity chromatography and analyzed by both the detection of Western blot and size exclusion chromatography;its antigen binding activity was examined by FACS and rosetting assay.Results:The data of DNA sequence showed that the anti CD3/anti CD20 Diabody was corrected.The Diabody was recovered in high yield(up to 1 mg/ml) after E taq purification and predominantly(90%) as a dimer.The Diabody binded Jurkat cells(CD3 +) and Daudi cells (CD20 +),respectively.Furthermore,the Diabody was capable of simultaneous binding to Jurkat cells and Daudi cells as shown by cellular rosetting.Conclusion:The anti CD3/anti CD20 BsF(ab') 2 was first recast into the Diabody format and succeeded to obtain high level expression.The results of some biological activity experiments indicated that the Diabody could bind to Jurkat cells and Daudi cells.
