肌旋毛虫免疫亲和层析抗原的制备及其特性

The Preparation and Characterization of Immunoaffinity Chro- matographic Antigens from Trichinella Spiralis in Muscles

以Sepharose 4B为载体,兔抗旋毛虫多克隆抗体为配体,纯化了旋毛虫的S_3抗原,得到亲和层析抗原.本实验结果表明,存在于S_3抗原中的主要抗原成分,同样存在于亲和层析抗原中,亲和层析抗原的分子量范围在103000～40000 u之间,等电点范围在pH4.7～8.8之间.亲和层析抗原经转印后,在硝酸纤维素膜上至少有7条显色带,其中分子量为48000 u,50000u,58000u和87000u的4种蛋白的抗原性较强,而以分子量为48000u的蛋白抗原性最强.

The immunoaffinity chromatographic antigens were prepared from S3 antigens with Sepharose 4B column containing rabbit polyclonal antibodies against Trichinella spiralis. Their biochemical and immunological properties were analysed on SDS-PAGE, slab gel isoelec-tric forcusing and Western blotting techniques. The results showed that the main components ofS3 antigens were also existed in immunoaffinity chromatographic antigen. Their molecular weights were from 40000 to 103000 daltons, and isoelectric points were from pH4.7 to pH8.8. There were at least 7 clear bands of protein on nitrocellulose exhibited by Western blotting technique. Among them the antigenicities of the proteins whose MW were 48000, 50000, 58000 and 87000 daltons were stronger and the first one strongest. When the antigens were used ,to detect trichinosis of pigs by ELISA, they were superior to Ss antigens in sensitivity and specificity.