Zhang Zhi-Chao, Hu Xue-Jun, Yang Qing
State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116012, China.
Hepatobiliary Pancreat Dis Int. 2004 Feb;3(1):77-81.
A single-chain antibody (ScFv) phage display library was created by cloning antigen-binding regions of V(H) (variable domain) and V(L) gene repertoires as fusion proteins with a minor coat protein of filamentous phage, from which high affinity completely humanized ScFv against PreS1 of hepatitis B virus could be screened and characterized.
A combinatorial library of phage-display human ScFv genes, which were derived from peripheral blood lymphocytes immunized by peptide PreS1 in vitro, was constructed. The library contained 7 x 10(8) clones.
After 3 rounds panning, a high affinity (K=10(-7)-10(-8)mol/L) ScFv specific to PreS1 was obtained. Sequence analysis showed that the V(H) belonged to the V(H)4 family and V(lambda) to V(lambda)4.
The described ScFv may provide a more satisfactory therapy. This application further illustrates that the method of in vitro antigen stimulation is expeditious for the source of human immune antibody library.
通过将V(H)（可变区）和V(L)基因库的抗原结合区作为与丝状噬菌体的次要外壳蛋白的融合蛋白进行克隆，创建了一个单链抗体（ScFv）噬菌体展示文库，从中可以筛选和鉴定出针对乙型肝炎病毒PreS1的高亲和力完全人源化ScFv。
构建了一个噬菌体展示人ScFv基因的组合文库，这些基因来源于体外经PreS1肽免疫的外周血淋巴细胞。该文库包含7×10(8)个克隆。
经过3轮淘选，获得了对PreS1具有高亲和力（K=10(-7)-10(-8)mol/L）的ScFv。序列分析表明，V(H)属于V(H)4家族，V(λ)属于V(λ)4。
所述ScFv可能提供更令人满意的治疗方法。该应用进一步说明体外抗原刺激方法对于人免疫抗体库的来源是快速有效的。
