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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
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- Authors
- Issue Date
- 2001
- Publisher
- Springer
- Citation
- Journal of Microbiology, vol.39 no.1, pp. 49-55
- Keywords
- 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 λ-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 λ-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.
- ISSN
- 1225-8873
- Language
- English
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