Preparation And Identification Of An Anti-Idiotypic Antibody Against Mcyst-LR

Microcystin-LR (short for MC-LR,MC) is a class of natural toxins generated by blue-green algaea. Scientists study suggests that the microcystin of drinking water and aquatic products has brought a great threat to the human health.The most common now known and content in a relatively and toxicity of larger is the number of species such as MC-LR, MC-RR, MC-YR. Then People take a bath, swimming and other water of recreation and sport, Skin contact the water which containing algal toxins can cause sensitive parts irritation such as eyes and skin. A small amount of drinking can cause acute gastroenteritis, Long-term consumption is likely to cause hepatocellular carcinoma. In order to detect the content of microcystin in water and food, the toxin in water and food be detect width, distance from the piie.Then Superimpose the effective of several single piles to simulate the effective of pile group. inspection.The advantage of prepare anti-idiotypic antibodies with Polyclonal Antibody technology is simpler and easier to operate than monoclonal antibody production technology, if it is great in demand, we need to prepare many times181.The main method is to use the monoclonal antibody (Abi)direct immunological test animals, and then extract and purification the serum. Frist of all,the poly-antibodies against MCYST-LR were purified by saturated ammonium sulfate, and then further purified by affinity chromatography with protein GThe result of SDS-PAGE showed that the MCYST-LR poly-antibodies were purified by affinity chromatography with protein G is effective.The yield of it was87.57%.The purified poly-antibodies were purified digested by papain under mild conditions. The antibody fragments were separated by DEAE-52ion-exchange chromatography to obtain the F(ab’)2fragments.The immunocompetence and yield of F(ab’)2fragments were measured. The results showed that the fragments immunocompetence reduced50%with1:12000.But it still can be used as Antigen to preparated anti-idiotypic antibody. We are used the fragments as new immunogen to immunited mouse of BalB/C by general immunited method and obtain the anti-idiotypic antibody against MCYST-LR.The new antibody was purified by the same method as saturated ammonium sulfate and affinity chromatography with protein G and measured the purify effect and concentration of antibody. The potency, specific and the best work of new antibody was measured by detection technology of ELISA.The result of SDS-PAGE showed that the purified effect effective and the concentration of IgG was27.1mg/mL, the best work of antigen is0.25ppm, the best work of antibody is1:1600.