抗-HBs单克隆抗体细胞系与酶联免疫吸附试验“一步法”检测HBsAg

ESTABLISHMENT OF THE CELL LINES SECRETING ANTIHBs MONOCLONAL ANTIBODY AND DETERMINATION OF HBsAg USING ELISA "ONE-STEP" TECHNIQUE

本文利用杂交瘤技术,经脾内直接注射免疫BALB/c小鼠,与骨髓瘤细胞(Sp 2/0)进行融合,建立了6株抗HEsMcAb,其中5株已能稳定地分泌单克隆抗体,并已得到腹水;5据中4株为抗a,1株为抗d,利用其中2株分别作为包被、酶标抗体,建立了酶联免疫吸附试验(ELISA)“一步法”检测HBsAg技术,经验证,该方法简便、快速、灵敏、特异、稳定。

A procedure is described for the production of HBsAg-specific monoclonal antibodies, which invo-lves immunization of BALB/c mice by intrasplenic injection with purified HBsAg and fusion of lymp-hocyte from spleen with SP2/0 mycloma line. Six positive hybrids were obtained, five of which havegot stable secreting ability throngh which mouse ascitic fluids were prepared. Four strains are specificfor the 'a' determinant of HBsAg. One is anti-'d' determinant. Two strains of anti-HBs McAb were used for conjugation with peroxidase and for coating thesolid phase respectively in the ELISA one-step system. The procedure was performed in a single hour'sncubation. It is found to be a rapid, sensitive, specific and reliable way to detect HBsAg.