Title Molecular Cloning and Characterization of cDNA Encoding Immunoglobulin Heavy and Light chain Variable Regions from Four Chicken Monoclonal Antibodies Specific to Surface Antigens of Intestinal Parasite, Eimeria acervulina
Author Ki Duk Song1, Jae Yong Han1, Wongi Min2, Hyun S. Lillehoj2, Sung Won Kim3, and Jin-Kyoo Kim3*
Address 1School of Agricultural Biotechnology, Seoul National University, Suwon 441-744, Korea; 2Parasite Biology, Epidemiology, Systemic Laboratory, Animal and Natural Resources Institute, Building 1040, BARC-East, USDA-ARS, Beltsville, MD 20705, USA.; 3Department of Microbiology, College of Natural Sciences, Changwon National University, Changwon 641-773, Korea
Bibliography Journal of Microbiology, 39(1),49-55, 2001,
Key Words Eimeria acervulina, chicken monoclonal antibody, gene conversion, antibody engineering, complementarity determining regions, somatic hypermutation
Abstract We have developed four chicken hybridomas secreting monoclonal antibodies to induce a protective immune response against the chicken disease avian coccidiosis, caused by the intestinal parasite Eimeria acervulina. However, since the amount of antibodies secreted from these hybridomas is too low or sometimes they lost their ability to produce antibodies, the hybridoma method is not satisfactory in the production of large amounts of chicken monoclonal antibodies. To bypass these problems, we applied the antibody engineering technology using polymerase chain reaction. We cloned and determined the sequences of variable domains of the four chicken monoclonal antibodies, namely, 2-1, 5D11, 13C8 and 8C3. The sequences comparison to germline sequences showed that the gene conversion mechanism might contribute to developing diversification of heavy and l-light chains in chicken antibodies. Several pseudogene families regarded as donors in gene conversion were identified at each framework region and the complementarity determining region of l-light chains. In addition, as expected, numerous changes of nucleotide sequences such as nucleotide substitution, insertion and deletion were found predominantly in complementarity determining regions, which are likely to be somatic hypermutations as a result of affinity maturation in antibody-producing cells.
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