only in the high capacity of the antibody library screening to high-affinity antibodies
In general, the affinity of the antibody capacity and the screened positive correlation only in the high capacity of the antibody library screening to high-affinity antibodies. Ribosome display technology as a new antibody library technology, its greatest superiority is to enable the storage capacity of 1013 ~ 1015, this is because it remove the transformation and cloning steps, completely in vitro, without relying on cells conversion and amplification, thus making the display library of larger capacity, more efficient screening, but also save time and effort, is an entirely in vitro expression and screening of functional proteins. In this study, the improvement of the two fragments overlap extension PCR method efficiently the antibody heavy chain and light chain are randomly connected together, also will serve as the intergenic regions CK gene fragments with 3 'end of the single-chain antibody gene, and then another to set up a pair of primers ribosome display in vitro expression of the T7 promoter and ribosome binding sites (Kozak sequence) into the connection good antibody gene fragments, so that was successfully constructed ribosome display the required template. The end of the spacer region to remove the stop codon after the completion of such protein to form the ribosome - the protein-mRNA complexes, which can be very convenient in vitro screening.