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Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27~(Kipl)immunized and non-immunized mice. After only one round of selection with rP27~(Kipl), clones from each library were chosen randomly and digested by Taq I and Hinf I. 11 of 64 clones from the immunized animal had consistent restriction pattern, while none of the 64 clones from the non-immunized animal had, except that one had the same fragments pattern as that of the 11 clones. The 12 fragments were expressed in E. coli BL21(DE3)/pET-28b(+) system. ELISA showed that some of the fragments could bind to rP27~(Kipl) specifically. All these results implied that specific antibody can be obtained by genetic engineering without hybridoma or many rounds of growth and panning selection.
Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27 ~ (Kip1) immunized and non-immunized mice. After only one round of selection with rP27 ~ (Kip1), clones from each library were chosen randomly and digested by Taq I and Hinf I. 11 of 64 clones from the immunized animal had consistent restriction pattern, while none of the 64 clones from the non-immunized animal had, except that one had the same fragment patterns as that of the 11 clones. The 12 fragments were expressed in E. coli BL21 (DE3) / pET-28b (+) system. ELISA showed that some of the fragments could bind to rP27 ~ (Kipl) specifically. All these results implied that specific antibody can be obtained by genetic engineering without hybridoma or many rounds of growth and panning selection.