Yasir Ashrif
Production, Purification And Concentration Of Rabbit Anti Goat I&G Antibodies - 2010
Antibodies are not only important in medical research but these are also important in treatment. In this study, the production, purification and concentration of polyclonal immunoglobulin G (IgG) antibodies against goat IgG immunoglobulins in rabbits was carried out. Partial purification of goat IgG obtained at 33 % ammonium sulphate saturation was 2.43 mg/mL. It was followed by Diethylaminoethyl (DEAE) cellulose ion exchange chromatography which gave purified fraction of IgG. Then it was concentrated by polyethylene glycol (PEG) and now the IgG concentration was found 3.17 mg/mL. After purification, different doses of IgG in combination with adjuvant were injected into nine, 8 months old rabbits. After immunization of rabbits, the blood samples were collected and antigoat IgG was purified as described above, this rabbit antigoat IgG concentration after purification was found 3.26 mg/mL. Production of these anti-IgG antibodies were tested by agar gel precipitation test (AGPT) and radial immunoassay. The titer of AGPT with goat and rabbit serum was 256 and the titer with IgG was 32. The purity of IgG was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), by obtaining 50 KDa bands of IgG heavy chains and 25 KDa bands of light chains. However, this purified rabbit anti-goat IgG when conjugated with horse radish peroxidase can serve to diagnose various microbial infections of goat through ELISA.
Institute of Biochemistry & Biotechnology
1197,T
Production, Purification And Concentration Of Rabbit Anti Goat I&G Antibodies - 2010
Antibodies are not only important in medical research but these are also important in treatment. In this study, the production, purification and concentration of polyclonal immunoglobulin G (IgG) antibodies against goat IgG immunoglobulins in rabbits was carried out. Partial purification of goat IgG obtained at 33 % ammonium sulphate saturation was 2.43 mg/mL. It was followed by Diethylaminoethyl (DEAE) cellulose ion exchange chromatography which gave purified fraction of IgG. Then it was concentrated by polyethylene glycol (PEG) and now the IgG concentration was found 3.17 mg/mL. After purification, different doses of IgG in combination with adjuvant were injected into nine, 8 months old rabbits. After immunization of rabbits, the blood samples were collected and antigoat IgG was purified as described above, this rabbit antigoat IgG concentration after purification was found 3.26 mg/mL. Production of these anti-IgG antibodies were tested by agar gel precipitation test (AGPT) and radial immunoassay. The titer of AGPT with goat and rabbit serum was 256 and the titer with IgG was 32. The purity of IgG was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), by obtaining 50 KDa bands of IgG heavy chains and 25 KDa bands of light chains. However, this purified rabbit anti-goat IgG when conjugated with horse radish peroxidase can serve to diagnose various microbial infections of goat through ELISA.
Institute of Biochemistry & Biotechnology
1197,T