ProImmune REVEAL & ProVE® Rapid Epitope Discovery System

Pro5® MHC Class I Pentamers

ProVE® Pentamers in Human Herpesvirus research

ProVE® Pentamers in Cancer Vaccine development

Application Notes: multiple staining with unlabeled Pentamers

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ProVE® MHC Class I Pentamer Libraries

ProVE® MHC Class I Pentamer Libraries provide a quick and cost effective way of screening a number of peptides implicated as epitopes in the immune response under investigation.

ProVE® Libraries bridge the gap between the screening of high numbers of T cell epitopes and the in-depth monitoring and characterization of single specificity CD8+ T cell immune responses with our individually manufactured Pro5® MHC Class I Pentamers. A cost reduction of more than 80% is achievable compared to the synthesis of standard MHC reagents.

 

ProVE® Pentamer Library Features
Rapid high throughput synthesis process	Fast delivery time (2-3 weeks). A large number of Pentamers can be delivered together.
Flexibility in supplying the peptide	Custom peptides can be synthesized by ProImmune or supplied by the customer.
Full QC for each Pentamer	Consistent and reliable performance compared with self-made tetramers or self-loaded dimeric MHC products.
Low price per multimer and no custom set up charges	Up to 80% cost reduction per reagent when compared to the synthesis of standard Pentamers

Product Specification

A ProVE® Pentamer Library is a set of custom Pentamers. The customer specifies the MHC allele under investigation and provides a small quantity of each of the peptides of interest. Alternatively, a custom synthesis of the chosen peptides may be ordered from ProImmune at competitive rates.

ProImmune manufactures the Pentamers using a rapid, high throughput, parallel synthesis process and supplies them unlabeled and at a minimum quantity of 20 tests. There is a minimum order of 10 Pentamers per ProVE® Library per allele.

ProVE® Pentamers are suitable for multiplexed staining of antigen-specific T cells in flow cytometry. This enables the user to identify and quantitate different populations of single antigen-specific CD8+ T cells very rapidly and attain conclusive validation of new T cell epitopes.

Quality Control

Each ProVE® Pentamer is QC tested. The protein concentration is determined and translated into quantity for each product. The customer pays for the complete synthesis run and receives those ProVE® Pentamers that pass quality control. The average QC pass rate for HLA-A*02:01 ProVE® Pentamers is 85% for peptides with scores >21 in the SYFPEITHI epitope prediction algorithm (www.syfpeithi.com).

Before proceeding with an order, ProImmune will review the sequences of the peptides submitted for inclusion in the ProVE® Pentamers and will notify the customer of any anticipated problem sequences.

Feature Comparison

Comparative Staining of PBMC with ProVE® MHC Pentamers and Pro5® MHC Pentamers

Peripheral blood lymphocytes were obtained from a healthy donor previously shown to respond to the A2-restricted CMV epitope, at approx. 0.15% of total PBMC. The left plot shows cells stained with the ProVE® MHC Pentamer, while the right plot shows cells stained with Pro5® MHC Pentamer made by the conventional process. The antigen-specific population is circled in red on both plots.

(Experimental procedure: cells were incubated with 1ug recombinant A2/CMV complex, followed by 1 test of PE-labeled Pro5® Fluorotag & 1 test of FITC-labeled anti-CD8 antibody (Clone LT8). Approximately 30,000 events are shown in each plot).