摘要
为探索利用噬菌体抗体库技术不经免疫制备抗体,在已有的抗乙肝病毒表面抗原(HBs)人Fab段基因的基础上,通过PCR引入突变,对其VHCDR3中12个氨基酸残基(95~100J)进行了完全随机化,对94和100K位残基进行了有限随机化,构建了半合成噬菌体抗体库,库容为2×108,58%含有随机化CDR3序列,用无关抗原小鼠IgG对该抗体库进行筛选,获得了三个可与小鼠IgG结合而不与HBs反应的克隆,经测定其CDR3的DNA序列,发现这三个克隆具有相同的CDR3,证明单纯改变VHCDR3可以得到新的特异性抗体,从而绕过免疫制备抗体。
In order to prepare antibodies by passing immunization we constructed a semisynthetic phage antibody library with randomized CDR3.Using anti HBs human Fab gene as starting material,12 amino acid residues in V H CDR3(95 ̄100J) were totally randomized and residues 94 and 100K were limited randomized by PCR.A phage antibody library of 2×10 8 with 58% of the clones containing randomized CDR3 was obtained.Panning of the library by unrelated Ag,mouse IgG,yielded 3 clones that could bind to mouse IgG but not HBs.Sequence analysis revealed that the 3 clones contained identical CDR3 sequence indicating they were derived from specific enrichment during panning,thus CDR3 randmization was feasible for preparing antibodies by passing immunization.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1997年第6期449-452,共4页
Chinese Journal of Microbiology and Immunology
基金
国家863计划项目
国家自然科学基金