Production of antipeptide antibodies
journal contributionposted on 29.04.2011, 00:00 by Bao-Shiang Lee, Jin-Sheng Huang, G.D. Lasanthi, P. Jayathilaka, Syed S. Lateef, Shalini Gupta
Peptides (8-20 residues) are as effective as proteins in raising antibodies, both polyconal and monoclonal with a titer above 20,000 easily achievable. A successful antipeptide antibody production depends on several factors such as peptide sequence selection, peptide synthesis, peptide-carrier protein conjugation, the choice of the host animal, and antibody purification. Peptide sequence selection is likely the most difficult and critical step in the development of antipeptide antibodies. Although the format for designing peptide antigens is not precise, several guidelines can help maximize the likelihood of producing high quality antipeptide antibodies. Typically, 5-20 mgs of peptide is enough for raising an antibody, for preparing a peptide affinity column, and for antibody titer determination using an Enzyme-Linked ImmunoSorbent Assay (ELISA). Usually, it takes three months to raise a polyclonal antipeptide antibody from a rabbit that yields ~90 mL of serum which translates into approximately 8-10 mgs of the specific antibody after peptide affinity purification.