title: 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.
authors: Protein Research Laboratory, Research Resources Center, University of Illinois at Chicago, 835 S. Wolcott Avenue, Chicago, IL, 60612, USA. boblee@uic.edu.
authors: Bao-Shiang
authors: BS
authors: Lee
authors: Protein Research Laboratory, Research Resources Center, University of Illinois at Chicago, 835 S. Wolcott Avenue, Chicago, IL, 60612, USA.
authors: Jin-Sheng
authors: JS
authors: Huang
authors: Protein Research Laboratory, Research Resources Center, University of Illinois at Chicago, 835 S. Wolcott Avenue, Chicago, IL, 60612, USA.
authors: Lasanthi P
authors: LP
authors: Jayathilaka
authors: Department of Chemical Engineering, Columbia University, 500 W. 120th Street, New York, NY, 10027, USA.
authors: Jenny
authors: J
authors: Lee
authors: Protein Research Laboratory, Research Resources Center, University of Illinois at Chicago, 835 S. Wolcott Avenue, Chicago, IL, 60612, USA.
authors: Shalini
authors: S
authors: Gupta
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D045163:Antibodies, Phospho-Specific / Q000096:biosynthesis* / Q000302:isolation & purification; D000918:Antibody Specificity; D000941:Antigens / Q000737:chemistry* / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D002352:Carrier Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry* / Q000276:immunology; D006433:Hemocyanins / Q000737:chemistry / Q000276:immunology; D057927:Hydrophobic and Hydrophilic Interactions; D010455:Peptides / Q000008:administration & dosage / Q000138:chemical synthesis* / Q000276:immunology; D011817:Rabbits; D012984:Software
chemical_list: D000911:Antibodies, Monoclonal; D045163:Antibodies, Phospho-Specific; D000941:Antigens; D002352:Carrier Proteins; D018985:Epitopes, B-Lymphocyte; D010455:Peptides; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin
keywords: Antibody titer; Antigenicity; Antipeptide antibody; Immunogenicity; Keyhole limpet hemocyanin (KLH); Peptide synthesis; Peptide–carrier protein conjugation; Phospho-specific antibodies; Polyclonal and monoclonal antibodies