REVEAL & ProVE® T Cell Epitope Discovery System | ProImmune

REVEAL & ProVE® Rapid Epitope Discovery System

REVEAL & ProVE® is a novel technology for analyzing proteins for potential T cell epitopes within only four weeks. The process saves weeks or months when compared with existing labor-intensive and expensive methods for epitope discovery.

The technology is widely applicable across many disease areas including all areas of cancer and infectious diseases. New CD4+ and CD8+ T cell epitopes identified with the help of this technology can be used as core building blocks for vaccine development or as targets for new immunotherapy.

Class I Epitope Discovery

Class II Epitope Discovery

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GEN Technical Article

Background to epitope discovery

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Class I REVEAL & ProVE® combines a powerful in vitro MHC-peptide binding assay with the rapid and cost effective synthesis of ProVE® MHC Class I Pentamers, ready for conclusive confirmation of epitopes on patient samples.

For a wide range of 56 HLA class II alleles (26 DR, 11 DQ, 19 DP), the REVEAL™ MHC-peptide binding assay and stability assays are available for rapid prediction of CD4+ T cell antigens in any protein sequence.

The REVEAL & ProVE® Custom Service

Protein or peptide sequences are chosen for investigation and the required modules of the custom service are selected from the following options:

 

Class I REVEAL & ProVE®

Class II REVEAL™
Step 1

PEPscreen®: Custom Peptide Synthesis by ProImmune, or peptides supplied by the customer

PEPscreen®: Custom Peptide Library Synthesis by ProImmune, plus optional set of peptides for cellular assays
Step 2

REVEAL™ MHC-Peptide Binding Assay for alleles:

A*01:01, A*02:01, A*03:01, A*11:01, A*24:02, A*29:02

B*07:02, B*08:01, B*14:02, B*15:01, B*27:05, B*35:01, B*40:01

H-2Kb, H-2Db, H-2Kd, H-2Dd, H-2Ld

Mamu-A*01, Mamu-A*02

REVEAL™ MHC-Peptide Binding Assay

26 DR, 11 DQ alleles and 19 DP alleles

List DR, DQ and DP Alleles and

Mouse H-2 IAb and IAd alleles

Step 3

ProVE® MHC Class I Pentamer synthesis for validation of CD8+ T cell epitopes in flow cytometry

Functional cellular assays with well defined REVEAL™ hit epitopes

Functional cellular assays with well defined REVEAL™ hit epitopes
Step 4

REVEAL™ on- and off-rate assays: full kinetic data for selected peptides

REVEAL™ on- and off-rate assays:

Full kinetic data for selected peptides for alleles DR1,2,3,4, 5 and 7

REVEAL™ Stability Assays for all DR, DQ and DP alleles

The customer receives a technical report with full data from the particular REVEAL™ assays selected, and in the case of the Class I service, all ProVE® Pentamers that pass quality control, for conclusive validation of new CD8+ T cell epitopes by flow cytometry.

Applications for REVEAL & ProVE® Epitope Discovery System

  • Rapid, comprehensive epitope mapping

  • Study of the immunogenicity of different variants of a protein, e.g. comparison of the variable and conserved regions of immunogenic proteins in different clades of a virus

  • Optimization of clinical monitoring e.g. selection of the optimal Pentamer specificities for monitoring a target patient group.

  • Identification of cross presentation of a peptide by several alleles

  • Improvement in vaccine design and efficacy

  • Mutagenesis studies: epitope binding optimization for therapeutic application, such as the insertion of anchor residues in peptide sequences with only partial or weak anchors

 

Advantages of the REVEAL & ProVE® Epitope Discovery System

Rapid

REVEAL™ binding assay determines MHC restriction of 94 peptides plus 2 controls in only five working days and may be carried out with several alleles in parallel. Class I ProVE® Pentamers can be produced in a further five working days for peptides generating positive results in the binding assay.

Informative

For Class I alleles, on- and off-rate binding assays may be carried out for specific peptides of interest in order to improve understanding of their kinetics.

Convenient

REVEAL & ProVE® is provided as a custom service. Assays are carried out in a controlled environment to ensure maximum accuracy and reproducibility.  No need for assay set up by the researcher, or purchase of reagents or software for analysis of results.

Flexible

Any starting set of peptides may be used, as long as the length matches the restriction of the alleles to be investigated. Peptides are synthesized by ProImmune for convenience, or may be supplied by the customer (for class I alleles only). The customer may select any or all of the modules that make up the REVEAL & ProVE® service. On- and off-rate binding assays may be ordered for class I alleles at a later date should they be required.

Affordable

REVEAL & ProVE® has been developed as an integrated system in order to offer rapid epitope discovery at an affordable price. When compared with existing labor-intensive methods and other commercial products REVEAL & ProVE® technology saves time and money, delivering results in only four weeks.

Summary

  • The only fully integrated, commercially available rapid epitope discovery system

  • Determines and confirms new T cell epitopes in weeks rather than months

  • Saves time and money when compared with traditional epitope discovery methods and other commercial products

  • ProImmune's custom service delivers fast and accurate results, removing the labor-intensive epitope discovery process and allowing the customer to focus on other important aspects of the research project

Improved Intellectual Property Position

Identification of the individual epitopes of a therapeutic protein allows separation of the intellectual property (IP) of that protein from the IP around the method of delivery or its therapeutic use.  During vaccine development, defining IP for specific epitopes increases the options for future changes in product design. If the IP for the specific antigens in a protein of interest is not secured, competitive organizations may have the opportunity to develop an obstructive patent position.

 

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