QuickSwitch™ Custom Tetramer Kits

Fast, High-Quality Generation of New Tetramer Specificities

Rapid, high-quality creation of custom Class I and Class II MHC tetramers in your own lab is now a reality! Create new specificity tetramers in just a few hours with QuickSwitch™, a proprietary technology for exchanging peptides on an MHC tetramer. With this kit, you will receive MHC allele with an irrelevant peptide bound. The addition of a peptide exchange factor, and your peptide of interest, results in the creation of your custom MHC tetramer ready to use- all in one day. The QuickSwitch™ Quant Tetramer Kit can quantify the exchange with the new peptide, thereby gathering binding affinity information for the MHC/peptide complex. QuickSwitch™ can be used to generate custom MHC tetramers, or to screen numerous peptides. This kit can be used for purposes such as epitope discovery, neoantigen vaccine research, verification of T cell staining using the new peptide specific tetramer, and more.

Functional screening of peptides for MHC class I binding is essential for vaccine design and immune monitoring. The QuickSwitch™ assay kit allows discrimination of MHC binding from non-binding peptides. This is particularly essential for the screening of immunogenic peptides from infectious agents or cancer neoantigens. Tetramers resulting from peptide exchange with selected peptides can then be used for immune monitoring. QuickSwitch™ kits are optimized for up to 10 peptide exchanges and multiple tests per resulting tetramer.

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QuickSwitch™ Class I Custom Tetramer Kit

Peptide exchange, quantification, cell staining, and flow cytometry analysis can all be performed in one day!

Steps for custom Class I MHC Tetramer creation*

*Total assay time is dependent on allele version of QuickSwitch™ kit.

High Throughput Immunogenic Peptide Discovery and Validation in 90 min.*

• Validate MHC binding peptides from in silico selected list of candidate peptides
• Generate new specificity tetramers for immune monitoring
• Perform functional stability studies for MHC binding peptides
• Compare epitopes to rank better binders and perform epitope mapping

Class I Kits Available

QuickSwitch™ Class II Custom Tetramer Kit

Custom Class II MHC Tetramers: Peptide exchange, quantify, cell staining, and FCM analysis all in your own lab!

Steps for custom Class II MHC Tetramer creation

*Total assay time is dependent on allele version of QuickSwitch™ kit.

Class II Kits Available

Additional QuickSwitch™ data

QuickSwitch™ HLA-A*02:01 tetramers detect similar percentages of low and high affinity CMV responses in PBMCs as classically folded tetramers

QuickSwitch™ Quant Kit can be used to assess peptide exchange so that you can select peptides with appropriate affinities to make functional tetramers prior to cell staining. In a study where HLA-A*02:01 QuickSwitch™ tetramer was incubated for 4 hours with two Mart-1 related peptides at a final of 20 μM in presence of peptide exchange factor #1, peptide exchange correlated with the theoretical peptide affinity of each peptide towards HLA-A*02:01.

QuickSwitch™ Quant Kit components:

• Tetramer with an irrelevant exchangeable peptide in the MHC groove
• A peptide exchange factor for catalyzing the peptide exchange reaction
• A high affinity MHC-binding reference peptide used as peptide exchange positive control
• FITC conjugated antibody specific of the exiting peptide
• Magnetic beads conjugated with anti MHC antibody for tetramer capture
• An Assay Buffer for diluting reagents and washing steps

H2-Kb Peptide-exchanged tetramers perform similarly to classically folded tetramers

H-2 Kb TRP2 used a negative control (#TB-5004-2; green), classically folded H-2 Kb OVA (#TB-5001-2; blue), and H-2 Kb QuickSwitch™ OVA (red) tetramer staining.

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QuickSwitch™ HLA-A*24:02 tetramers perform similarly to classically folded tetramers

Data show CD3+ PBMCs stained with classically folded tetramers (A,E) or with QuickSwitch™ tetramers obtained by peptide exchange with the HLA-A*24:02 CMV peptide (B,C,D) or the HIV negative control peptide (F,G,H). 2 x 105 cells in 50 μL PBS-BSA-NaN3 buffer were stained for 30 min at RT with 1 μL of anti CD3-PC5.5 mAb (clone OKT3), 1 μL anti CD8-FITC (clone RPA-T8) (MBL) and 0.25 μg tetramer. Cells were fixed with a 0.5% formaldehyde PBS solution. Cells were analyzed on a Cytoflex S flow cytometer (Beckman Coulter). Cell doublets were discriminated using SSC-W/SSC-A gating.

MHC Quant Monomer Kit

See Product TB-7300-KM-10

Create custom MHC Tetramer from MHC Monomers

MBL International’s MHC Quant Monomer Kit allows you to create your own custom MHC Tetramer from MHC Monomers. You may already know your peptide binds well with the MHC allele or you may want to screen some peptides to determine peptide-MHC binding. This MHC Quant Monomer kit comes with MHC Monomer bound with an irrelevant peptide. Perform peptide exchange on the monomer with your peptides of interest and quantitate the exchange. Then, tetramerize your new MHC Monomers bound with your peptide(s) of interest and add your desired conjugate. The resulting custom MHC Tetramer can be used to detect antigen specific T cells. MBL International’s revolutionary MHC Tetramer technology continues to provide unique products for solutions to advance T cell research.

MBL International continuously updates and adds new alleles to our current offerings. If you do not see your allele of interest, please contact us to inquire for more information.

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Citations

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