Preparation of Monoclonal Antibodies against Major Outer Membrane Protein of Neisseria Gonor-rhoeae / 中华皮肤科杂志

ABSTRACT

Objective To establish a rapid,sensitive and specific diagnostic test for detecting Neisse-ria gonorrhoea.Methods The major outer membrane proteins(P Ⅰ )in different gonococcal serogroups were obtained by isolation of outer membrane complex with CTB-ethanol precipitation,the outer membrane proteins were extracted with Z 3,14 and EDTA,and purified with DEAE-Sepharose CL-6B to obtain P Ⅰ .Hybridoma cell lines producing McAbs against P Ⅰ were established with lymphocyte hybridoma techniques.Results The molecular weight of P Ⅰ A and P Ⅰ B were determined with SDS-PAGE as35.2kDa and36.7kDa,respectively.Five hybridoma cell lines producing McAbs continuouslly and stably against P Ⅰ A and P Ⅰ B were obtained,in-cluding two hybridoma cell lines producing McAbs against P Ⅰ A and three hybridoma cell lines producing McAbs against P Ⅰ B.The titers of McAbs in the supernatants in the cultures and in abdominal ascites of BALB/c were from1:64to1:256and from1:4096to1:16384,respectively;and the specificity of the McAbs against P Ⅰ A and P Ⅰ B was so high that they easily reacted with N.gonorrhoeae but did not with other antigens such as N.meningitidis etc.Conclusion The purified P Ⅰ and the McAbs obtained in the study provide a basis to establish a rapid,sensitive and specific diagnostic test for detecting N.gonorrhoea.