抗宋内氏志贺氏菌脂多糖单克隆抗体的制备及其特性鉴定

Preparation and Identification of Monoclonal Antibodies against Lipopolysaccharide of Shigella sonnei

本文用宋内氏志贺氏菌加热致死菌体免疫BALB/c小鼠,纯化脂多糖筛选,获得2株抗宋内氏菌LPS的杂交瘤细胞株。制备了小鼠腹水抗体,并对杂交瘤细胞株进行了鉴定。2株McAb的Ig类型为IgG2、IgG2a。交叉试验表明,2株McAb与痢疾志贺氏菌、福氏志贺氏菌、侵袭性大肠杆菌、亲水气单胞菌、鼠伤寒沙门氏菌以及类志贺毗邻单胞菌O5、O15、O26、O39血清型无交叉反应。

Monoclonal antibodies ( McAbs ) were prepared by fusing SP2/O cells with spleen cells of BALB/c mice immunized with the antigen of Shigella sonnei. Antibodies in hybrid culture supernatant were detected by ELISA and positive hybrids were cloned using limiting dilutions. The number of chromosomes of two hybridoma cell lines (2E6, 5Bl2) ranged from 91 to 105. The immunoglobulin class/subclass of McAbs 2E6 and 5Bl2 was IgG3 and IgG2a, respectively. Titration of two McAbs (2E6> SBl2) using ELISA showed that their titers in the ascites reached 1:6400000 and 1:3200000, respectively. After repeated frozen storage and revival, the two hybridoma cell lines were capable of steady production of high titers of McAbs. 11F was used to investigate the cross-reaction of the McAbs and other enterobacteria and the results showed that McAbs demonstrated no cross reaction with other tested enterobacteria.