Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27kip1-immunized and non-immunized mice. After only one round of selection with rP27kip1, clones from each library were ch...Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27kip1-immunized and non-immunized mice. After only one round of selection with rP27kip1, clones from each library were chosen randomly and digested by Tag 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 BL2l(DE3)/pET-28b( + ) system. ELISA showed that some of the fragments could bind to rP27kip1 specifically. All these results implied that specific antibody can be obtained by genetic engineering without hybridoma or many rounds of growth and panning selection.展开更多
文摘Through mRNA extract, RT and a series of PCR, phage antibody libraries were constructed from rP27kip1-immunized and non-immunized mice. After only one round of selection with rP27kip1, clones from each library were chosen randomly and digested by Tag 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 BL2l(DE3)/pET-28b( + ) system. ELISA showed that some of the fragments could bind to rP27kip1 specifically. All these results implied that specific antibody can be obtained by genetic engineering without hybridoma or many rounds of growth and panning selection.
