利用在体外构建的三肽囊素抗独特型抗体可变区VH和VL基因,通过重叠延伸反应(SOE),以(Gly4Ser)3为连接肽,将VH和VL基因连接成为VH-Linker-VL ScFv,且ScFv DNA与噬菌粒载体pHENI的连接产物转化于大肠杆菌TGl,经辅助噬菌体M13K07感染后,获得重组的鸡源抗三肽囊素抗独特型抗体全套单链噬菌体抗体库,并将该抗体库展示在噬菌体表面,以利用噬菌体展示技术的强大筛选能力筛选出与三肽囊素同功能单链抗体(Bursin-ScFv)。
The repertoire of VH and VL gene fragments was amplified from cDNA templates. ScFv was constructed by splicing by over.lap extension reaction using (Gly4Ser)3 as a linker. The ScFV DNA was inserted into phagemid vector (pHENI) and transfected into E, coli TGI that was infected with helper phage MI3K07. A ScFv phage display library of the Bursin anti-idiotype antibody was successfully constructed.