Anti-Keratin Human Antibody: Gene Cloning, Expression And Affinity Maturation

Numerous investigations have showed that anti-keratin auto antibody (AK auto Ab) is a very important element in human immune system. In some pathological states such as psoriasis and contact dermatitis, a certain serum level of the antibody could inhibit the progression of the diseases, and is beneficial for the recovery from the diseases. Studies both in vitro and in vive animal models indicated that anti-keratin antibodies have therapeutic potential for such disorders. For serotherapy, monoclonal antibodies of human origin are preferred. But human hybridomas are difficult to make and require human immunization. Recent advances in the field of molecular immunology have enabled the isolation of human monoclonal antibodies more efficiently and more easily by engineering and expressing antibody fragments in E.coli using phage display. In the present study we explored the use of the genetic engineering technique to generate anti-keratin human antibodies with good specificity and high affinity from phage antibody library.Semisynthetic phage antibody library of 4.0 × 10~8 members was previously constructed from semi-synthetic V genes consisting of human genomic V segments and synthetic CDR3 fragments. Panning of the library against human epidermal keratin was performed. Phages eluted from each selection were used to infect E.coli TG1 cells and...