PepSpots™

Peptide Arrays for Mapping Protein Interactions

JPT Peptide Technologies developed a well validated technology (PepSpot™) to provide state-of-the-art B-cell epitope mapping in a reliable and speedy manner. The comprehensive all-in-one service includes: the cost-efficient generation of peptide scans spanning the entire sequences of antigens on membranes or on glass chips and the incubation of these membranes and chips with antibodies for detection (epitope mapping).

Mapped epitopes can be functionally characterized by rapid determination of key residues playing a definite role in the binding process. In addition, the method is well suited to detect de novo epitopes for antibodies with unknown antigens by employing JPT's complex random peptide libaries.

The customer will receive a comprehensive project report consisting of detailed descriptions of all experiments made, all resulting data and a comparision of the detected binding sites with the corresponding crystal structure of the antigen protein (if available).

More than 100 scientific papers describing the application of this technology and the hundreds of successful mapping projects performed at JPT over the last decade distinguish the PepSpot™ technology as the method of choice for fast and reliable antibody epitope mapping.

Applications for PepSpots™

• Antibody epitope mapping
• Functional characterization of mapped epitopes
• Characterization of protein-protein contact sites:
• Receptor-ligand
• Enzyme-substrate
• Protein modules involved in signal transduction processes
• Recognition sequences for heat shock proteins

Functional characterization of epitopes by substitutional analyses

Benefits of the PepSpots™ peptide membranes

• Rapid, economical, and accurate synthesis using robotics
• Reusable membranes
• Hydrophilic surface of cellulose membranes
• Minimizes unspecific hydrophobic interactions with the target molecule
• Easy detection using standard ELISA protocols

Take advantage of our comprehensive All-In-One-Service

• Synthesis of PepSpot™ peptides
• Detection with your protein and data interpretation
• Functional characterization of mapped epitopes
• Complete and confidential documentation of results

References:

• Peptide arrays on cellulose support: SPOT synthesis, a time and cost efficient method for synthesis of large numbers of peptides in a parallel and addressable fashion, Hilpert et al., Nature Protocols Vol. 2 (2007), p. 1333 - 1349 (abstract)

• Structure-based epitope and PEGylation sites mapping of phenylalanine ammonia-lyase for enzyme substitution treatment of phenylketonuria, Gamez et al., Mol. Genet. Metab., 2007 (abstract)

• Analysis of Antibody Response in Humans to the Type A OspC Loop 5 Domain and Assessment of the Potential Utility of the Loop 5 Epitope in Lyme Disease Vaccine Development, Buckles et al., Clin. Vaccine Immunol, Vol. 13(10), 2006, p. 1162-1165 (abstract)

• Identification of distinct antibody epitopes and mimotopes from a peptide array of 5520 randomly generated sequences, Reineke et al., J. Immun. Methods Vol. 267 (2002), p. 37-51(abstract)

• Peptide arrays: from macro to micro, Reimer et al., Curr. Opin. Biotechnol. Vol. 13 (2002), p. 315-320 (abstract)

• Applications of peptide arrays prepared by the SPOT-technology, Reineke et al., Curr. Opin. Biotechnol. Vol. 12 (2001), p. 59-64 ( abstract)

• Coherent Membrane Supports for Parallel Microsynthesis and Screening of Bioactive Peptides, Wenschuh et al., Biopolymers Vol. 55 (2000), p. 188-206 ( abstract)

• Antigen sequence and library-based mapping of linear and discontinuous protein-protein-interaction sites by spot synthesis, Reineke et al., Curr. Top. Microbiol. Immunol. Vol. 243 (1999), p. 23-36 (abstract)

• Molecular Basis for the Binding Promiscuity of an Anti-p24 (HIV-1) Monoclonal Antibody, Kramer et al., Cell Vol. 91 (1997), p. 799-809 (abstract)