Production of different antibodies after simultaneous immunization of animals with two antigens.
We propose a method of simultaneous immunization with two different antigens for isolation of two types of antibodies from the same antiserum. Bacterial proteins (Bacillus stearothermophilus glyceraldehyde-3-phosphate dehydrogenase and Escherichia coli GroEL chaperonin) served as the antigens. Affinity purification of antibodies was carried out using two columns: with covalently immobilized glyceraldehyde-3-phosphate dehydrogenase or GroEL chaperonin. During stage I, the antiserum was applied onto the column with immobilized glyceraldehyde-3-phosphate dehydrogenase, after which antibodies to glyceraldehyde-3-phosphate dehydrogenase were eluted. During the next stage, the antiserum without antibodies to glyceraldehyde-3-phosphate dehydrogenase was passed through the column with immobilized GroEL and antibodies to chaperonin were isolated. Antibodies to glyceraldehyde-3-phosphate dehydrogenase and to GroEL had high titers and exhibited no cross-reaction.
Pleten'
A P
AP
A. N. Belozyorskii Institute of Physico-chemical Biology, M. V. Lomonosov Moscow State University. pleten@belozersky.msu.ru
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000917:Antibody Formation; D018834:Chaperonin 60 / Q000276:immunology*; D003429:Cross Reactions; D004926:Escherichia coli / Q000737:chemistry; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000276:immunology*; D007114:Immunization / Q000379:methods*; D011817:Rabbits
D000906:Antibodies; D018834:Chaperonin 60; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases