adr和adw亚型HBsAg单克隆抗体的制备

Preparation and of monoclonal antibody against hepatitis B surface antigen of subtypes adr and adw

目的制备adr和adw亚型乙型肝炎表面抗原(hepatitis B surface antigen,HBsAg)单克隆抗体,并用其区分两亚型HBsAg。方法利用杂交瘤细胞融合技术制备adr和adw亚型HBsAg单克隆抗体,采用间接ELISA法进行筛选;优化鉴别试验抗原包被剂量,并对HBsAg亚型鉴别试验进行验证。结果共获得抗两亚型HBsAg的单克隆抗体7株;鉴别试验抗原的最佳包被剂量为10μg;1H6和4D3株单抗检测2份编盲adw亚型HBsAg样品的adwA450/adrA450值均大于2,判定其为adw亚型;1E9株单抗检测2份编盲adr亚型HBsAg样品adrA450/adwA450值均大于2,判定其为adr亚型;3A5株单抗检测4份编盲样品的adrA450/adwA450值和adwA450/adrA450值均小于2,未能区分两亚型HBsAg。结论抗adr亚型单抗1E9株及抗adw亚型单抗1H6株和4D3株可用于adr亚型汉逊酵母乙肝疫苗研制过程中HBsAg亚型的鉴别。

Objective To prepare the monoclonal antibodies （McAb） against hepatitis B surface antigen （HBsAg） of subtypes adr and adw, and use for differentiation of the two subtypes. Methods The McAbs against HBsAg of subtypes adr and adw were prepared by hybridoma cell fusion technique, and screened by indirect ELISA. The dosage for coating of antigen for identity test was optimized, and the identity test for HBsAg subtype was verified. Results Seven hybridoma cell strain secreting McAbs against HBsAg of subtypes adr and adw were obtained. The optimal dosage for coating of antigen for identity test was 10μg. The adwA450/adrA450 values of two blinded HBsAg subtype adw samples determined with McAbs secreted by strains 1H6 and 4D3 were more than 2, indicating the McAbs as subtype adw. The adrA450/adwA450 values of two blinded HBsAg subtype adr samples determined with MeAb secreted by strain 1E9 was more than 2, indicating the McAb as subtype adr. However, the adrA450/adwA450 and adwA450/adrA450 values of four blinded HBsAg samples determined with McAb secreted by strain 3A5 were less than 2, indicating that McAb failed to identify the HBsAg of two subtypes. Conclusion The McAb against HBsAg of subtype adr secreted by strain 1E9, and those against subtype adw secreted by strains 1H6 and 4D3 may be used for the identification of HBsAg subtype during development of Hansenula anomaladerived hepatitis B vaccine of subtype adr.