Epitope Mapping Service Using Mimotope Variation Analysis (MVA) Technology
Study 1. Defining of the minimal epitope sequence (Positional Weight Matrix) in peptide antigen binding using MVA.
Epitope mapping of two monoclonal antibodies raised against identical nonapeptide YPYDVPDYA showed that the binding of these antibodies to the immunogen is different. For mAb1, amino acids 1,2,3,4, and 7 are crucial for mAb binding, whereas for mAb2, the recognized epitope consists of amino acids 1, 3, 4, and 6. In the visualized minimal epitope sequence profiles aligned with the immunogen, the consensus sequence of the epitope is delineated such that the overall height of the amino acid representation letter indicates the relative frequency of the specific amino acid at this position.
Study 2. Multi-epitope mapping using MVA.
Rabbit polyclonal antibodies were generated using peptide immunogen APQPGLASPDSPHDPCK. Epitope mapping using standard MVA protocol identified four partially overlapping epitopes (A, B, C, D).
In the visualized minimal epitope sequence profiles aligned to the immunogen, the consensus sequence of the epitope is delineated such that the overall height of the amino acid representation letter indicates the relative frequency of the specific amino acid at this position. A, B, C, D – epitopes in the immunogen sequence.
Advantages of MVA Based Epitope Mapping:
- Precise – Identifies and distinguishes all, including overlapping, epitopes.
- Quantitative and qualitative – Epitope description includes Positional Weight Matrix of amino acid residues in the peptide epitope motif.
- Universal – No need for prior knowledge of the immunogen.
- Cost effective – Saves money compared to competing techniques, such as peptide array.
- Epitope mapping defines the binding specificity of an antibody. Protobios’s Mimotope Variation Analysis (MVA) technology gives a detailed description of epitopes and mimotopes and has several advantages over other technologies.
- In a standard MVA Epitope mapping protocol, three million antibody-captured peptides from the peptide display library will be analyzed and clustered. In general, monoclonal antibody analysis will result in one epitope, whereas polyclonal antibody analysis may result in numerous epitopes.
- The immunogen competition assay (ICA) is a recommended procedure for confirming the epitope-specific binding activity of antibodies. The ICA provides means to discriminate between true and non-specific (crossreactive) targets for the antibody.
BASIC CONCEPT OF MVA
- Ig CAPTURE
- AFFINITY SELECTION USING PEPTIDE DISPLAY LIBRARY
- NEXT GENERATION SEQUENCING
- DATA ANALYSIS
In MVA epitope mapping service we deliver:
- Epitope sequences
- Ranking of epitopes
- Positional Weight Matrix of amino acid residues in the peptide epitope
- Alignment of epitopes on antigen
- Competition profile (if required)
- For more information please contact our technical service by
Phone +49 (0) 551 707 2270 or E-mail.