Development and characterization of a recombinant chicken single-chain Fv antibody detecting Eimeria acervulina sporozoite antigen.
Chicken monoclonal antibody (mAb), 8C3, which is reactive with a sporozoite antigen of Eimeria acervulina, is a potential therapeutic agent against avian coccidiosis caused by Eimeria spp. However, production of large amounts of 8C3 mAb in cell culture system is labor intensive and not cost-effective. Accordingly, recombinant single chain variable fragment (ScFv) antibody was constructed by amplification of the V(H) and V(L) genes from chicken hybridoma, 8C3 and when expressed in E. coli gave 5 mg l(-1). The expressed protein showed antigen binding activity equivalent to that of the parental mAb. In addition, nucleotide sequence comparison of 8C3 gene to the germline chicken V(L) genes suggested that the gene conversion with (V)lambda pseudogenes might contribute to the diversification of V(L) genes in chickens.
Park
Kyung Je
KJ
Department of Microbiology, College of Natural Sciences, Changwon National University, Changwon 641-773, Korea.
Park
Dong Woon
DW
Kim
Chun Hee
CH
Han
Beom Ku
BK
Park
Tae Sub
TS
Han
Jae Yong
JY
Lillehoj
Hyun Soon
HS
Kim
Jin-Kyoo
JK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods*; D002645:Chickens; D003048:Coccidiosis / Q000517:prevention & control; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004306:Dose-Response Relationship, Immunologic; D004539:Eimeria / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000378:metabolism; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D007135:Immunoglobulin Variable Region / Q000737:chemistry; D007167:Immunotherapy / Q000379:methods; D008957:Models, Genetic; D008969:Molecular Sequence Data; D011201:Poultry Diseases / Q000517:prevention & control; D011994:Recombinant Proteins / Q000737:chemistry*; D012689:Sequence Homology, Nucleic Acid; D034101:Sporozoites / Q000378:metabolism
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D018076:DNA, Complementary; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins