马拉硫磷抗性致倦库蚊酯酶单克隆抗体的制备及鉴定

PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST ESTERASE OF MALATHION RESISTANT CULEX QUINQUEFASCIATUS

用马拉硫磷抗性致倦库蚊酯酶免疫经ＰＰＤ刺激的小鼠制备了５株单克隆抗体，经鉴定均为ＩｇＧ１，各株单抗两两相加的相加指数（Ａ．Ｉ．）均未超过５０％。酯酶与１∶１０００稀释的腹水孵育１ｈ后即失去分解底物活性。免疫印迹结果显示，单克隆抗体识别的抗原在８个群体（或品系）均有一６４ｋＤａ蛋白带，反应强度及蛋白带数量与抗性程度有关。单抗对幼虫及苄呋菊酯抗性品系酯酶基本无反应。

Five clones of monoclonal antibodies (McAbs) were generated from BALB/c mice which were activized with PPD and immunized with esterases extracted from malathion resistant Culex quinquefasciatus . All 5 clones of McAbs belonged to the IgG 1 subtype, additive index (A.I.) of every two McAbs did not exceeded 50%. After a l h of incubation, the McAbs at the titer of 1∶1 000 completely inhibited the activity of the amplified esterases. Of 8 strains of adult mosquitoes studied by Western blot assay, all exhibited the 64 kDa esterase band recognized by the McAbs, six (SP,FS,SH,CD,RD,RM) showed more bands of different MW (43-60 kDa), the higher the organophosphorus resistant level, the more and the denser the bands. The above McAbs had little or no effect against the esterases of dimethrin resistant strain and the larva stage of malathion resistant Culex quinquefasciatus .