用亲和层析技术鉴别布氏菌强弱毒差异抗原成分

采用CDI和CCA两种方法活化Sepharose4B,制备亲和层析柱,并对这两种方法的优劣进行了比较。分别用超声液破碎和Ribi机器压榨制备布氏菌强毒M28株和疫苗M25株的可溶性抗原,作为亲和层析柱的配基。抗M28的血清经过M28和M5柱,洗脱液中的抗体再分别和M28、M5抗原反应经电转印检查证实,M28抗原中的一条蛋白带只与M28柱洗脱液反应,不与M5柱洗脱液反应。这种只与M28柱洗脱液反应的蛋白可能即是布氏菌强毒M28株所特有的抗原成分,也就是强毒株和弱毒株的差异抗原成分。

Study of Identifying the Different Antigens of the wild-type Blucella Strain M28 to the Vaccine Strain Ms by Affinity Chromatography

We have identified the different antigens of the wild-type Blucella strain M28 to the vaccine strain M5 by affinity chromatography. As a ligand the antigens of the wild-type Blucilla strain M28 and the vaccine strain M5 were respectively coupled to sepharose 46 with 2, 4, 6-trichloro-5-triazinl carbonyl. The columns were eluteb with M propionic acid and 0.05M diethylamine.ELISA and western blotting were used assaying the results. In ELISA the reaction between M28 antigens and the elution of M28 column with propionic acid was stronger than that of Ms column. The diethyl-amine elutions of both columns were similar. In western blotting, the elution of M28 column had a band with M28 antigens. The band dispeared with M5 antigens. This band also dispeared in unboand solutions of both M28 and M 5 Columns. We believe the antigens in this band is a specific antigen of M28.