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.
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.
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.
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).