McCafferty J, Griffiths A D, Winter G, Chiswell D J
MRC Laboratory of Molecular Biology, Cambridge, UK.
Nature. 1990 Dec 6;348(6301):552-4. doi: 10.1038/348552a0.
New ways of making antibodies have recently been demonstrated using gene technology. Immunoglobulin variable (V) genes are amplified from hybridomas or B cells using the polymerase chain reaction, and cloned into expression vectors. Soluble antibody fragments secreted from bacteria are then screened for binding activities. Screening of V genes would, however, be revolutionized if they could be expressed on the surface of bacteriophage. Phage carrying V genes that encode binding activities could then be selected directly with antigen. Here we show that complete antibody V domains can be displayed on the surface of fd bacteriophage, that the phage bind specifically to antigen and that rare phage (one in a million) can be isolated after affinity chromatography.
最近已证明利用基因技术可制造抗体的新方法。使用聚合酶链反应从杂交瘤或B细胞中扩增免疫球蛋白可变(V)基因,并将其克隆到表达载体中。然后筛选细菌分泌的可溶性抗体片段的结合活性。然而,如果V基因能够在噬菌体表面表达,那么对V基因的筛选将会发生变革。携带编码结合活性的V基因的噬菌体随后可以直接用抗原进行筛选。在此我们表明,完整的抗体V结构域可以展示在fd噬菌体表面,噬菌体能够特异性结合抗原,并且经过亲和层析后可以分离出罕见的噬菌体(百万分之一)。