人工合成HLA-B_(27)多肽抗原的单克隆抗体研究

PREPERATION OF MONOCLONAL ANTIBODIES AGAINST CHEMICALLY SYNTHESIZED PEPTIDFS DERIVED FROM HLA-B 27.1 MOLECULE

本文报道用人工合成HLA-B_(27)抗原决定基的多肽为免疫原来制备McAb的研究.根据计算机对B_(27)分子一级结构的B细胞识别序列的分析结果,用固相法合成了B_(27)分子多态区抗原决定基上P_(62-24)与P_(69-84)两个肽段,分别与KLH和BSA载体偶联,免疫RALB/c小鼠.当按常规方法免疫动物进行5次融合,所得17株抗小肽McAb均可与载体分子有较强的交叉反应.当在融合前的末次加强免疫改为仅用不接载体的小肽来进行,三次融合就获得了9株不与载体分子发生交叉反应的特异性抗肽McAb,为今后鉴定出可和天然B_(27)分子起反应的McAb提供了基础.文中对这二种免疫方案还作了比较和讨论。

Based on the result of epitope analysis on the primary molecular structure of HLA-B27 through computer programs, two peptides corresponding to the amino acid sequences 63-84 and 69-84 of the molecule were synthesized by solid-phase method and coupled with either KLB or BSA carriers. These synthetic peptides were administered to BALB/c mice with two immunization protocols. After routine immunization protocol, we made fusions for 5 times and got 17 clones of McAbs reactive with the immunizing peptides, but also crossreactive significantly with carriers. Applying modified immunization protocal in which non-conjugated peptide was used in the final boost, after fusion for 3 times, we got 9 clones of McAbs reactive specifically to pepties without crossreaction with carriers. Our experimental results indicated that design of immunization protocals is very important for perperation of specific McAb against small peptides.