Construction Of A Large Human Phage-display Antibody Library And Screening Antibodies Against AIV

The therapeutic potential of monoclonal antibodies (mAb) was quickly realised after the hybridoma technique allowed their development in the mid 1970s. But the mouse anitibodies do not fit for the therapy because of HA1MA (human antimurine antibody response) which can lead a unexpectable immune reaction and make the antibodies degrade quickly in vivo. In the following years, the rapid development of human antibody technology make antibodies using as therapeutic drug to be possible. While techniques of humanization, human memorr B cell immortalization and antibody engineering booming, more and more antibodies became performing clinicle researches, and went into the market.With easily operating and high-throughput screening, phage display antibody library shows to be one of the most useful techniques for gain human antibodies. If the normal process using for construction the library, the capacity would be much smaller than it should be. It's for the reason that a large part of antibody genes would be lost in PCR, ligation and transfromation. In the other side, the antibody light chain and heavy chain genes in library can not recombine freely as in immune system. All of these make gain a good antigen especial reaction antibodies to be difficult. However, after presenting a recombination system, the diversity of library can be enlarged obviously. Additionally, a large library with 10" were gained.In this research, on the basis of the primers which could amplify almost the whole antibody V-region genes, a normal single-chain phage antibody library has been prepared successfully. Also the Cre/lox recombination system was established. This system ensure the diversity of our scFv library.Antibodies play important roles in protection against and recovery from virus infection. Techniques of human antibody bring a sparking point for therapy of viral diseases. Herein, the scFv library was sCreened with different H5N1 AIV strains in liquid/solid-phase, while detection with ELISA and HAL Then the positive clones were expressed in E. coli. as scFv proteins. Some clones showed cross-activity to different H5 virus strains. Remarkably, one of them can greatly cross-react with H1, H3 and H5 virus. The results depict that the antibodies against conserved epitops in virus have been gotten. It appears an approachable method for developing therapeutic antibody to influenza viruses who usally change themselves.