Peptide-ready MHCs

Quick and simple in-house peptide loading for custom MHC monomers or tetramers

What are prMHCs?

KACTUS peptide-ready MHCs (prMHC) are MHC monomers and tetramers absent of antigenic peptides. The prMHCs are stabilized and ready for loading the neoantigen peptide of your choosing. They are ideal for generating custom MHC peptide tetramers and high throughput peptide screening. The prMHCs are expressed from HEK293 cells and have >95% purity.

Do you offer a peptide-loading protocol?

KACTUS offers a quick and simple protocol for loading peptides in-house. Contact us to receive the protocol.

What can I do with a prMHC?

Assess MHC/TCR Binding Affinity

Enabling rapid and high-quality creation of custom Class I and II MHC tetramers, our Peptide-Ready MHCs (prMHC) provide a streamlined and user-friendly approach to developing new custom MHC monomers and tetramers in just a few minutes directly in your lab. Simply by mixing your peptide of interest with one of our prMHCs and incubating at room temperature, you generate a Peptide-MHC complex, which can be utilized for the creation of custom MHC tetramers and monomers or employed in high-throughput screening of peptides. This innovative system holds applications across various realms including epitope discovery, neoantigen vaccine research, and verification of T cell staining, among others.

Generate a custom MHC peptide monomer or tetramer

In neoantigen identification, assessing the immune functionality of the antigenic peptide is critical, which includes evaluating its binding affinity to MHC and its reactivity with TCR. To facilitate this, KACTUS has introduced a range of functional Peptide-Ready MHCs products. These can serve as a ready-to-use loading system to assist in loading antigen peptides and subsequently form a new, complete MHC peptide complex, thereby significantly aiding your neoantigen
research studies by ensuring efficiency and reliability.

High Throughput Peptide Screening

prMHCs are instrumental in the functional screening of peptides for MHC class I binding, a critical component in vaccine design and immune monitoring. They provide the distinct ability to discriminate between MHC binding and non-binding peptides, which is particularly pivotal when screening immunogenic peptides derived from infectious agents or cancer neoantigens. Subsequently, the generated Peptide-MHC complexes can be used for immune monitoring.

Class I Peptide-Ready MHC (prMHC).

Shop Alleles

Browse available alleles in our stabilized peptide-free form.

Other Alleles

KACTUS supports expression of other MHC alleles including class II alleles and non-human species. Submit our custom MHC form to request an allele not listed above.

Ordering Information

Order Online

To order a prMHC product online, browse available products and add to cart to pay online via credit card.

To pay via PO, send your order information to orders@kactusbio.us.

Custom MHC

Submit our custom MHC form to request a a custom pMHC or prMHC.

Request Quote

Contact us to request a standard or bulk quote, or for more information.

Browse All Available Products

Browse our selection of in-stock Peptide-Ready MHC Complexes.

Performance Validation of prMHCs

Figure 1. Demonstrated via ELISA assay, biotinylated Human Peptide Ready HLA-A*11:01&B2M Monomer loaded peptide (VVVGADGVGK) has a high affinity with HLA-A*11:01&B2M&KRAS G12D TCR. The EC50 is 110 ng/mL.

Figure 2. Demonstrated via SPR assay, biotinylated Human Peptide Ready HLA-A*11:01&B2M Monomer loaded with peptide (VVVGADGVGK) has a high affinity with HLA-A*11:01&B2M&KRAS G12D TCR. The affinity constant is 8.5 nM. 

Figure 3. Demonstrated via SPR assay, Human Peptide Ready HLA-A*02:01&B2M& Monomer loaded with peptide (FMNKFIYEI) has a good affinity with HLA-A*02:01&B2M&AFP TCR. The affinity constant is 0.32 µM.

Figure 4. Demonstrated via FACS assay, fluorescent-labeled Human Peptide-Ready HLA-A*02:01&B2M Tetramer loaded with peptide (SLLMWITQC) can bind with HLA-A*02:01&B2M&NY-ESO-1 TCR cells.

Stability Testing of KACTUS prMHCs

Freeze Thaw Stability

KRAS G12D peptide was added to biotinylated human HLA-A*11:01 and freeze thawed up to five times. Activity was analyzed via ELISA. Immobilized TCR was added to the plate at 2μg/mL (100μL/well). Results are a dose response curve for Biotinylated
Human KRASG12D (HLA-A*11:01), His Tag with EC50s of 0.22/0.16/0.18/0.15/0.15/19 μg/mL.

Biotinylated human HLA-A*11:01 was freeze-thawed up to five times after which KRAS G12D peptide was added. Activity was analyzed via ELISA. Immobilized TCR was added to the plate at 2μg/mL (100μL/well). Results are a dose response curve for Biotinylated Human KRASG12D (HLA-A*11:01), His Tag with EC50s of 0.21/0.13/0.18/0.16/0.16 μg/mL.

Stability Testing at 4C

KRAS G12D peptide was added to biotinylated human HLA-A*11:01 and incubated at 4℃ for 0, 7, 14, and 21 days. Activity was analyzed via ELISA. Immobilized TCR was added to the plate at 2µg/mL (100µL/well). Results show a dose-response curve for Biotinylated Human HLA-A*11:01&B2M&KRAS G12D (VVVGADGVGK) monomer, His Tag with EC50s of 0.19/0.11/0.13/0.14/0.16 µg/mL.

Biotinylated human HLA-A*11:01 was incubated at 4°C for 0, 7, 14, and 21 days, after which KRAS G12D peptide was added. Activity was analyzed via ELISA. Immobilized TCR was added to the plate at 2µg/mL (100µL/well) on the plate. Results show a dose-response curve for Biotinylated Human HLA-A*11:01&B2M&KRAS G12D (VVVGADGVGK) monomer, His tag with EC50s of 0.16/0.11/0.26/0.32/0.36 µg/mL.

Stability Testing at 37C

KRAS G12D peptide was added to biotinylated human HLA-A*11:01 and incubated at 37℃ for 0, 1, 3, and 5 days. Activity was analyzed via ELISA. Immobilized TCR was added to the plate at 2µg/mL (100µL/well). Results show a dose response curve for biotinylated human KRAS G12D (HLA-A*11:01), His tag with EC50s of 0.19/0.22/0.16/0.22/0.29/10.56 µg/mL.

Biotinylated human HLA-A*11:01 was incubated at 37℃ for 0, 1, 3, and 5 days, after which KRAS G12D peptide was added. Activity was analyzed via ELISA. Immobilized TCR was added to the plate at 2µg/mL (100µL/well). Results show a dose-response curve for Biotinylated Human KRAS G12D (HLA-A*11:01), His tag with EC50s of 0.19/0.22/0.52/0.46/0.43 µg/mL.

Neoantigen Peptides and Their Role in MHC/TCR Interactions

Neoantigens are highly specific targets and ideal targets for immunotherapy. Degraded neoantigen peptides can bind with major histocompatibility complex (MHC) molecules, forming complexes that are subsequently transported to the cell surface. These complexes are recognized by T-cell receptor (TCR), triggering an immune response. MHC polypeptide complexes constitute a category of neoantigen targets, and their distinctive interaction with TCR holds immense importance in the advancement of neoantigen-based immunotherapies, including TCR-T cell therapy, antibody drugs, and tumor vaccines.

Identification of the affinity of peptide loaded MHC with TCR [1].

Other MHC/TCR Products & Services

Custom MHC

Contact us to request a custom MHC with choice of allele, peptide, monomer/tetramer, etc.

Soluble TCR Expression

Custom expression of soluble TCR with TCR/MHC binding analysis via SPR.

MHC Peptide Monomers/Tetramers

Browse our catalog selection of MHC peptide complexes.

HLA-G and LILRs

Learn more about our full portfolio of HLA-G and LILRA/LILRB proteins.

prMHC FAQs

Ordering Information

We have a catalog selection of prMHCs in-stock and available to ship. Contact orders@kactusbio.us to place an order using a PO or pay via credit card on our website. We also offer custom prMHCs by submitting this form or contacting sales@kactusbio.us.

To order a custom prMHC, please submit this form or contact sales@kactusbio.us.

For prMHC catalog products, visit the specific product page to view the lead time. For custom prMHC orders, the lead time is 6 to 8 weeks. 

Our Products

KACTUS provides catalog and customized prMHC monomers, biotinylated monomers, and fluorescent tetramers. 

We analyze prMHC products for molecular weight (Tris-Bis PAGE), purity (Tris-Bis PAGE & SEC-HPLC), endotoxin, and concentration. Our prMHC products have > 95% purity and < 1 EU/µg.

Yes, KACTUS provides custom prMHC-I and prMHC-II for all available MHC alleles. Contact us at sales@kactusbio.us to request a custom MHC.

prMHCs can be stored for at least 12 months at -80°C. 

About Peptide-Ready MHCs

Peptide-ready MHCs (prMHC) is an MHC that doesn’t contain any pre-loaded peptide and can be loaded with custom peptides for TCR binding and cell staining. 

A prMHC monomer consists of an α-chain and β-2 microglobulin. It is a single subunit of prMHC tetramer.

It is a prMHC monomer with biotin at the C-terminal Avi tag.

A fluorescent prMHC tetramer is a complex of four prMHC monomers with fluorophores conjugated to each monomer subunit of the prMHC complex.

Expression of prMHCs

Yes, KACTUS prMHCs are a propriety design based on our SAMS™ Protein Engineering Platform.

Yes, prMHC are a single-chain design of α-chain and β-2 microglobulin developed by our protein engineering team.

Yes, prMHCs are purified from HEK293 cells.

prMHCs are affinity-purified via His tag at the C-terminus with additional size exclusion chromatography steps.

No, prMHC tetramerize in vivo during expression in HEK293, a KACTUS propriety design.

Fluorescent Labeling

We currently offer APC-equivalent, FITC-equivalent and PE conjugation for prMHC tetramers.

APC-equivalent is a 90 kDa protein, with excitation peak at 609 nm and emission peak at 643 nm.

No, APC-equivalent is an in vivo label, part of KACTUS proprietary single chain design for fluorescent prMHC tetramer. 

Protocols & Applications

Yes, contact support@kactusbio.us for a protocol.

Yes, KACTUS prMHCs can be used for peptide screening. 

Yes, contact support@kactusbio.us for a protocol.

Still have questions? Email support@kactusbio.us to get in touch.

References

[1] Moritz A, Anjanappa R, Wagner C, Bunk S, Hofmann M, Pszolla G, Saikia A, Garcia-Alai M, Meijers R, Rammensee HG, Springer S, Maurer D. High-throughput peptide-MHC complex generation and kinetic screenings of TCRs with peptide-receptive HLA-A*02:01 molecules. Sci Immunol. 2019 Jul 19;4(37):eaav0860.

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