3种抗糖皮质激素抗体的制备、纯化与鉴定

Preparation, Purification and Certification of Antibody to Corticosteroids

采用活性酯法合成地塞米松(DEX)、倍他米松(BET)、双氟米松(FLM)3种免疫原和包被原,用免疫原分别免疫2只新西兰大白兔,5次免疫后进行采血获得抗血清。用盐析与DEAE-纤维素离子交换联用的方法纯化抗血清,再用间接酶联免疫法测定抗血清的效价,其中较好的抗血清效价分别高达345600、614400、307200,具有较好的亲和性,可以满足抗原抗体反应的动力学要求。

Three immunogens and coating-antigens were prepared by conjugating derived dexamethasone, betamethasone and flumethasone with bovine serum albumin, ovalbumin respectively. Each immunogen was applied to immunize two New Zealand rabbits to obtain anti-serums. After five immunization, the antiserum was collcted from the blood, and purified by salt fractio combined with DEAE. The indirect ELISA was adopted to determine the efficiency of the purified antiserum. Among all the antiserum samples, the better efficiency of the antiserum could be as high as 345600, 614400 and 307200 respectively, which had sufficient affinity to meet the requirements of antigen-antibody reaction kinetics.