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Updated: Jul 7, 2026

Immunization of Alpacas (Lama pacos) with Protein Antigens and Production of Antigen-specific Single Domain Antibodies
Published on: January 26, 2019
Production of different antibodies after simultaneous immunization of animals with two antigens
1A. N. Belozyorskii Institute of Physico-chemical Biology, M. V. Lomonosov Moscow State University. pleten@belozersky.msu.ru
Abstract:
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.
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