Abstract: Objective To produce humanized anti-keratin antibodies with high affinity. Methods Panning of phage antibody libraries against human epidermal keratin was conducted to select specific antibodies. Random mutation method and chain-shuffling technique for the gene of the antibodies were employed to improve antibody affinity. The binding activity, specificity and affinity of the obtained antibodies were determined and analyzed. Results Four clones of either IgG or IgM anti-keratin antibodies were seleeted from semisynthetic or natural antibody libraries. One of them was seleeted to undergo affinity maturation. Random mutation method for the gene did not result in any improvement in antibody affinity. By chain-shuffling technique, however, affinity level was markedly increased and an antibody with affinity of 8×10¹⁰/M was isolated. Conclusion It is feasible to generate humanized antikeratin antibodies using genetical engineering technique.

Key words: Keratin, Antibodies, monoclonal, Antibody affinity, Gene expression