Natural human-derived ribosome display antibody library and identification of.doc

 PAGE \* MERGEFORMAT 10 Natural human-derived ribosome display antibody library and identification of [Abstract] Objective To construct a large storage capacity, diversity good ribosome display single chain antibody (ScFv) library, the basis for further screening ScFv. Method from peripheral blood lymphocytes of healthy volunteers were extracted mRNA, were amplified by RT PCR antibody heavy chain variable region (VH) and light chain variable region (VL) DNA, and then connected to form ScFv DNA. be connected into E.coli JM109 E. coli T Vector, white selection, picked 9 ScFv clones sequenced to identify the assembly. by in vitro transcription and translation of the antibody library were identified. Results VH, VL and ScFv DNA were about 350,650 and 1 100 bp. The test was successfully constructed ScFv ribosome display template. Conclusion Construction of large-capacity natural human derived ribosome display ScFv library for further screening by affinity enrichment to obtain various types of ScFv basis. [Keywords:] Antibodies, monoclonal, ribosome, Escherichia coli, gene library Single-chain antibody (Single Chain fragment variant, ScFv) through genetic engineering methods to the heavy chain variable region (VH), light chain variable region (VL) connection recombinant protein has a molecular weight than the full antibody is small, Higher permeability characteristics, is more suitable for drug targeting and immune biological treatment. ScFv can be constructed through the traditional hybridoma technology or phage display technology to prepare and come out in recent years, the ribosome display technology (ribosomal display, RD) is a species from cells transfected with the expression of other factors and extracellular show the complete system engineering [1], simplifying the ScFv screening process is expected to filter to the high affinity of the ScFv. In this study peripheral blood lymphocytes of healthy volunteers was materials to construct a large-capacity