Back to Search
Start Over
Antibody Production with Synthetic Peptides.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2016; Vol. 1474, pp. 25-47. - Publication Year :
- 2016
-
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.
- Subjects :
- Amino Acid Sequence
Animals
Antibodies, Monoclonal isolation & purification
Antibodies, Phospho-Specific isolation & purification
Antibody Specificity
Antigens immunology
B-Lymphocytes cytology
B-Lymphocytes immunology
Carrier Proteins administration & dosage
Carrier Proteins chemistry
Carrier Proteins immunology
Enzyme-Linked Immunosorbent Assay
Epitopes, B-Lymphocyte immunology
Hemocyanins chemistry
Hemocyanins immunology
Hydrophobic and Hydrophilic Interactions
Peptides administration & dosage
Peptides immunology
Rabbits
Software
Antibodies, Monoclonal biosynthesis
Antibodies, Phospho-Specific biosynthesis
Antigens chemistry
Epitopes, B-Lymphocyte chemistry
Peptides chemical synthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1474
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 27515072
- Full Text :
- https://doi.org/10.1007/978-1-4939-6352-2_2