Physico-chemical and antigenic characterization of unconventional heavy chain antibodies of Indian desert camel (Camelus dromedarius L.).
Heavy chain antibodies (HCAbs) of IgG2 and IgG3 subtypes were purified from the sera of Indian desert camel (Camelus dromedarius L.) by ammonium sulphate precipitation, followed by ion-exchange chromatography on DEAE-cellulose and affinity chromatography on protein A-sepharose and protein G-sepharose, and characterized by SDS-polyacrylamide gel electrophoresis, agar gel immunodiffusion (AGID), counter-immunoelectrophoresis (CIEP), immunoelectrophoresis (IEP), ELISA and immunoblotting. IgG2 and IgG3 were found to have molecular mass 46.77 kDa and 43.65 kDa, respectively by SDS-PAGE under reducing conditions. They migrated in beta-region in IEP and could be detected in CIEP, because of being more negatively charged and smaller size. Anti-camel IgG3 cross-reacted in AGID, ELISA and immunoblotting with IgGs of pig and ruminants (cattle, buffalo, sheep and goat), but not with immunoglobulins from horse, dog, guinea pigs, mice, fish, poultry and human. The present findings suggest close antigenic relationship of camels with pigs and ruminants.
Immunology Section, Department of Veterinary Microbiology, Ch. Charan Singh Haryana Agricultural University, Hisar 125 004, India.
Sharvan
S
Sehrawat
Ajit
A
Singh
D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D000941:Antigens / Q000737:chemistry*; D002162:Camelus; D002627:Chemistry, Physical / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry*; D013045:Species Specificity
D000941:Antigens; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D000645:Ammonium Sulfate