Custom Pro5^® MHC Class I Pentamers

ProImmune has over 10 years experience making custom MHC multimers specific to customers’ requirements. Our technical expertise and experience in making thousands of MHC-peptide complexes means that we are able to provide custom reagents efficiently and with a high rate of success.

Our customers can expect the same consistency and reliability from our custom reagents as our regularly ordered catalog Pentamers. In addition, all labeled custom Pro5® Pentamers come with a six month performance guarantee, and unlabeled custom Pentamers have a twelve month guarantee.

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You can now opt for faster dispatch by choosing the Rapid Synthesis option:

Rapid Custom Pro5^® MHC Class I Pentamer Service

When time is critical in your project, you can now choose custom Pentamers for dispatch in only 12 working days. This service, which includes peptide synthesis, is designed to beat the timescales of any external source of MHC multimers, such as tetramers - whether commercial or core facility.

Your rapid synthesis custom Pentamer is made to the same uncompromising quality standards as our standard delivery-time custom Pentamers and is supplied with the same guarantee, 6 months for a labeled Pentamer, 12 months for an unlabeled Pentamer.

Rapid Pentamer Service Specification

(1) The rapid synthesis service is limited to a maximum of 4 custom MHC Class I Pentamers per order and to the 50 test pack size.

(2) The rapid synthesis service is subject to availability at the time of ordering.

(3) Dispatch is 12 working days from the day that the order is accepted by ProImmune. An order is accepted if the custom peptide sequence submitted passes the criteria set by ProImmune for custom Pentamer synthesis, and all the relevant information is provided to allow processing of the order, e.g. full invoice and delivery addresses, method of payment, customer account not on hold. Following order acceptance and processing, email confirmation is sent to the customer with an expected dispatch date.

(4) Each rapid synthesis custom Pentamer is subject to a Rapid Synthesis Charge (product code FRSC), see price list for details

(5) The Rapid Synthesis Charge will only be made if the Pentamer is dispatched on or before the 12th working day following order acceptance, or if the Pentamer fails synthesis and quality control on or before the 12th working day following order acceptance.

(6) The Rapid Synthesis Charge will not be made if the Pentamer is dispatched after the 12th working day following order acceptance, or if the Pentamer fails synthesis and quality control after the 12th working day following order acceptance.

(7) We expect to meet our 12 day deadline in most cases. However, due to the nature of synthesis of new custom peptide sequences we cannot guarantee that all orders can be dispatched within 12 days. ProImmune strongly advises that experiments using custom items are not set up prior to arrival of the product with the end user.

(8) Delivery time is subject to the transit conditions and destination. Custom Pentamers are delivered from the UK by an express service overnight to Europe, in 1-2 days to USA & Canada and in 4 days to the rest of the world.

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Ordering Custom Pro5^® Pentamers and related products

Case Study: Epitope Discovery in Prostate Cancer

The Use of H-2Db / HCIRNKSVI (PSA 65-73) Custom Pro5® MHC Class I Pentamer in the Characterization of an Immunodominant CTL Epitope of PSA in Mice

Pavlenko, M. et al. (2005) Identification of an immunodominant H-2Db-restricted CTL epitope of human PSA. Prostate 64: 50-59 [PubMedID: 15651071].

Prostate cancer is a serious condition affecting 1 in 6 men. Prostate specific antigen (PSA) expression is increased in prostate cancer and so is exploited not only for diagnosing and monitoring prostate cancer, but also as a potential target for immunotherapy. By working with Custom Pro5^® Pentamers from ProImmune, Pavlenko et al. have identified and validated an immunodominant cytotoxic T lymphocyte (CTL) epitope of PSA in C57BL/6 mice. A combined bioinformatics approach using the SYFPEITHI website (www.syfpeithi.com), and biochemical MHC-peptide stabilization assays was used to define the candidate epitope (H-2Db / HCIRNKSVI) and a custom Pro5^® Pentamer was synthesized.

PSA-specific CTLs were induced by immunizing mice with a plasmid expressing PSA (pVax-PSA). By using both functional assays, intracellular cytokine staining and detection of PSA-specific CD8⁺ T cells with Pro5^® Pentamer staining, the authors were able to demonstrate correlating frequencies of both IFN-g positive and Pentamer positive T cells following in vitro stimulation with the specific peptide.

The authors conclude that “Pentamer technology enables detection of T cell receptor specific T-cell populations and allows, in combination with functional assays, the discrimination between anergy and tolerance induction during effector responses. H-2Db Pentamers assembled with this peptide are an efficient tool for the monitoring of PSA-specific CTL responses after DNA vaccination”.

Figure: 12 days after immunization as described, the splenocytes were restimulated for 5 days in vitro with 1nM of psa65-73 (HCIRNKSVI) peptide (A). The PSA-specific T cells were detected by staining with the H-2Db/HCIRNKSVI Pentamer (B). The splenocytes were stimulated for 5hr with 10nM of the psa65-73 or GP33 (negative control) peptides and stained with the antibodies against CD8 and IFN-gamma.