Identification and characterization of single chain antibody of cancer stem cell surface marker CD133 scFv CD133

VernacularTitle:肿瘤干细胞表面标志物CD133单链抗体基因的分离与鉴定
Author: Yu HAN ; Bingxi ZHOU ; Yanrui ZHANG ; Shuangyin HAN
Publication Type:Journal Article
Keywords: Tumor stem cells; Antibodies,monoclonal; Immunologic techniques
From: Cancer Research and Clinic 2011;23(4):230-233,246
CountryChina
Language:Chinese
Abstract: Objective To identify,construct and express scFv CD133,verify its biological function.Methods VL and VH were isolated from hybridoma of mAb CD133 by using antibody engineering technology.Its DNA sequencing and CDR were determined.scFv CD133 was then cloned into pET32a,transformed into Origami,induced by IPTG,purified by Ni2+-NTA His resin.Its affinity and specificity were tested by NH4SCN elution and ELISA.Results The size of VL and VH of scFv CD133 was 339 bp and 342 bp,which coded 113 and 114 amino acid separately.Its VL belonged to mouse Igκ chain and VH belonged to mouse IgG heavy chain subtype I.The molecular weight of scFv CD133 was about 27 × 103 which was testified by SDSPAGE and Western blot.Its affinity and specificity were also verified.Conclusion scFv CD133 has been successfully constructed and expressed in Origami,which could supply basis for target therapy of CD+133 cancer stem cell.