Antibody Production with Synthetic Peptides

Abstract

Peptides (usually 10-20 amino acid residues in length) can be used as effectively as proteins in raising antibodies producing both polyclonal and monoclonal antibodies routinely with titers higher than 20,000. Peptide antigens do not function as immunogens unless they are conjugated to proteins. Production of high quality antipeptide antibodies is dependent upon peptide sequence selection, the success of peptide synthesis, peptide-carrier protein conjugation, the humoral immune response in the host animal, the adjuvant used, the peptide dose administered, the injection method, and the purification of the antibody. Peptide sequence selection is probably the most critical step in the production of antipeptide antibodies. Although the process for designing peptide antigens is not exact, several guidelines and computational B-cell epitope prediction methods can help maximize the likelihood of producing antipeptide antibodies that recognize the protein. Antibodies raised by peptides have become essential tools in life science research. Virtually all phospho-specific antibodies are now produced using phosphopeptides as antigens. Typically, 5-20 mg of peptide is enough for antipeptide antibody production. It takes 3 months to produce a polyclonal antipeptide antibody in rabbits that yields ~100 mL of serum which corresponds to ~8-10 mg of the specific antibody after affinity purification using a peptide column.

Accumulation Profiles of Embryonic Salt-Soluble Proteins in Maize Hybrids and Parental Lines Indicate Matroclinous Inheritance: A Proteomic Analysis

“…Thus, we produced its antibody anti-GLB1 to further analyze its accumulation in the reciprocal hybrids and parental inbred lines. Generally, it is sufficient to elicit an effective immunization response by using a 10–20 amino acid peptide (Wu et al, 2013 ; Lee et al, 2016 ). The immunoreactivity of the polyclonal antibody anti-GLB1 was examined using SDS-PAGE, 2-DE, and immunoblot analyses (Figure S2 ).…”

Duplication and diversification of trehalase confers evolutionary advantages on lepidopteran insects

“…Three peptides corresponding to BmTreh1a (FQAILKNPRRAAHWGYMAEQ), BmTreh1b (GRKVRPDLIQEEDC), and BmTreh2 (CGSYDKARKDYFVNR) were selected, synthesized, and used as antigens to generate rabbit polyclonal antibody according to a standard protocol (Lee, Huang, Jayathilaka, Lee, & Gupta, ). To rule out cross‐reactivity between primary antibodies (anti‐BmTreh1a, anti‐BmTreh1b and anti‐BmTreh2), we performed Western blotting assay to detect antibody specificity using purified BmTreh1a, BmTreh1b, and BmTreh2 proteins.…”

“…and BmTreh2 (CGSYDKARKDYFVNR) were selected, synthesized, and used as antigens to generate rabbit polyclonal antibody according to a standard protocol (Lee, Huang, Jayathilaka, Lee, & Gupta, 2016). To rule out cross-reactivity between primary antibodies (anti-BmTreh1a, anti-BmTreh1b and anti-BmTreh2), we performed…”

Can Immunization of Hens Provide Oral-Based Therapeutics against COVID-19?

“…Therefore, one of the strategies in the development of protective antibodies against SARS-CoV-2 is to know the contact regions between the Spike protein and ACE2 and to generate antibodies against these critical epitopes of this protein in the hope that these antibodies will block the entry of the virus and prevent the development of the disease [82]. Peptide-based immunization in birds would guarantee the safety of this particular type of vaccination, not aimed at disease prevention, but at the mass production of polyclonal antibodies [83][84][85].…”