Shrivastav Tulsidas G, Basu Anupam, Kariya Kiran P
Department of Reproductive Biomedicine, National Institute of Health and Family Welfare, Munirka, New Delhi, India.
J Immunoassay Immunochem. 2003;24(2):191-203. doi: 10.1081/IAS-120020084.
The first step of enzyme linked immunosorbent assay (ELISA), namely, adsorption of antigen or antibody to the plastic microtiter well plate, was studied as a function of insolubility of IgG in water. Immobilization efficiency was assessed in terms of number of wells coated per milliliter of primary antiserum. We have compared different coating/immobilization protocols, i.e., direct and indirect immobilization of primary antibody to the plastic microtiter well plate using carbonate buffer and phosphate buffer with glutaraldehyde. We have observed efficient coating when the immobilization of primary antibody through an immunobridge technique was performed, where water was used as a coating medium. It gave a higher number of wells coated per milliliter of anti-serum (primary or secondary) than other compared coating protocols and it allowed the use of serum (non-immune) and anti-serum (primary and secondary antibody) dilutions, avoiding the need for gamma-globulin purification from normal and immunized serum.
酶联免疫吸附测定（ELISA）的第一步，即将抗原或抗体吸附到塑料微量滴定板上，作为IgG在水中不溶性的函数进行了研究。根据每毫升抗血清包被的孔数评估固定化效率。我们比较了不同的包被/固定化方案，即使用碳酸盐缓冲液和含戊二醛的磷酸盐缓冲液将一抗直接和间接固定到塑料微量滴定板上。我们观察到，当通过免疫桥技术进行一抗固定时，使用水作为包被介质可实现高效包被。与其他比较的包被方案相比，每毫升抗血清（一抗或二抗）包被的孔数更多，并且允许使用血清（非免疫血清）和抗血清（一抗和二抗）稀释液，无需从正常血清和免疫血清中纯化γ-球蛋白。
