沙门氏菌鞭毛蛋白属共同抗原表位的定位

用淋巴细胞杂交瘤技术研制成４株沙门氏菌属特异单抗ＣＢ８，ｄｅ７，ｃｃ６，ＤＤ４，在免疫转印试验中，证实它们所识别的抗原表位位于鞭毛蛋白上，ＥＬＩＳＡ相加试验及竞争试验显示，ＣＢ８和ｃｃ６与ｄｅ７，ＤＤ４识别的抗原表位不同，表明鞭毛蛋白的属共同抗原具有多种表位的特性。用胰蛋白酶消化鞭毛蛋白，ＳＤＳ－ＰＡＧＥ结果出现两条新带，分子量分别为４２０００（Ｆ４２），２７０００（Ｆ２７），长时间消化，最终只剩下Ｆ２７条带，它可抵制该酶的消化。对该酶切样品的免疫转印结果表明，沙门氏菌鞭毛蛋白属共同抗原表位出现于Ｆ４２和Ｆ２７上，而相应的沙门氏菌分型Ｈ抗原单抗或因子血清识别的表位也在Ｆ４２和Ｆ２７上都得到了证实。这为沙门氏菌鞭毛蛋白抗原的多样性及结构和功能研究提供了证据。

LOCATION OF MULTIPLE EPITOPES OF COMMIN ANTIGEN ON SALMONELLA FLAGELLINS WITH MONOCLONAL ANTIBODIES

Western blotting(WB)test revealed that CB8,de7,cc6, DD4 were reacting with antigenic determi-nants that were common to all Salmonella flagellins studied, respectively,Added enzyme-linked immunosorbent assay(ELISA) and competitive ELISA provided evidence that there were multiple epitopes of common antigen on the flag-ellin molecules,these epitopes were not related to serotypic antigens.Limited proteolysls of flagellins by trypsin resultsin a degradation pattern,The terminal regions of flagellins are very sensitive to proteolysis.These parts are degradedinto small oligopeptldes at the very early stage of a mild digestion that yields a relatively stabe fragment with a molecu-lar weight of 42 000(F42),further proteolytic degradation results in a stable 27000(F27)Mr fragment;WB test re-vealed that CB8,de7,cc6,DD4 were reacting with F42 and F27,corresponderingly,serotypic H antigen monoclonalantibody was reacting with F42 and F27 also,These results supply some new acknowledgement and evidence forSalmonella antigens,structure and functions of parts of Salmonella flagellins,respectively,moreover,they have agood use for studying evolutlon of Salmonella.