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目的 制备亲和力高、性能良好的人源性抗角蛋白抗体。方法 以人表皮角蛋白为抗原从噬菌体抗体库中克隆特异性抗角蛋白抗体,采用基因随机点突变和链替换(chain-shuffing)技术对所获抗体进行亲和力成熟,并对其生物学性能进行一系列鉴定。结果 经过筛选半合成抗体库和天然抗体库,成功获得了4株能与角蛋白特异性结合的人源性抗角蛋白IgG或IgM抗体;对抗体基因的随机突变未产生预期的性能改进,又选择其中活性最好的一株IgG抗体,通过轻链替换和重链替换使抗体亲和力得到了有效提高,达到8×1010/M的理想水平。结论 用基因工程的方法可以制备高质量的人源性抗角蛋白抗体。
Objective To prepare human anti-keratin antibodies with high affinity and good performance. Methods The specific anti-keratin antibody was cloned from the phage antibody library using human epidermal keratin as antigen. Affinity maturation of the obtained antibody was carried out by gene point mutation and chain-shuffing techniques. The biological performance of the anti- A series of identification. Results Four strains of human anti-keratin IgG or IgM antibodies that specifically bound to keratin were successfully obtained after screening the semi-synthetic antibody library and natural antibody library. Random mutation of the antibody gene did not produce the expected performance improvement. One of the most active IgG antibodies was selected and the antibody affinity was effectively increased by light chain substitution and heavy chain replacement, reaching the ideal level of 8 × 1010 / M. Conclusion High-quality human anti-keratin antibodies can be prepared by genetic engineering.