Services – NewPureMHC

Services Overview

Pure MHC offers its suite of proprietary tools, expertise, and resources to support therapeutic development efforts in oncology, autoimmunity and infectious disease indications on a contract research basis. Contact us to discuss your project needs today.

MHC Immunopeptidomics Survey

Pure MHC provides comprehensive liquid chromatography-mass spectrometry (LCMS) analysis of the naturally occurring MHC immunopeptidome on virtually any biological input sample. Pure MHC has extensive experience purifying MHC class I, MHC class II, and non-classical MHC class I from a wide variety of biological samples such as cell lines, primary cells and tissues, blood plasma, etc. Using state-of-the-art LCMS equipment and techniques, Pure MHC will survey the immunopeptidome to discover peptide ligands de novo with high levels of sensitivity, generating a comprehensive analysis of the immunopeptidome from biological samples. Pure MHC can also provide semi-quantitation on immunopeptidome datasets enabling comparative studies.

Validation of HLA-A2 Target Presentation

Pure MHC screens your HLA-A2 restricted peptide targets against one or all three of Pure MHC’s proprietary HLA-A2 immunopeptidome databases. This analysis can be used to validate existing HLA-A2 targets and/or screen for peptides that may cross-react with HLA/peptide therapeutics (CAR-T, TCR-T, TCR mimics).

Pure MHC Healthy Immunopeptidome Database and Repository (PHIDR):

HLA-A2 immonopeptidome from normal tissues.
Covers more than 40 distinct tissues from throughout the human body.
Every donor carefully screened to exclude any malignancies or chronic viral infections. Each tissue snap-frozen at the time of organ recovery with very low post-mortem intervals (3-14 hours, average 9 hours).

Primary Tumor Database:

HLA-A2 immunopeptidome from HLA-A2 positive primary tumor samples.
60 total tumor samples from colorectal carcinoma (CRC), non-small cell lung carcinoma
(NSCLC), and ovarian carcinoma (OC).

Tumor Cell Line Database:

HLA-A2 immunopeptidome from a variety of colorectal, lung and ovarian tumor cell lines.

Study Antigen Presentation by Macrophages Using THP-1 Model

THP-1 is a human leukemia monocytic cell line that is commonly used as a model to study monocyte/macrophage functions, signaling pathways, and antigen presentation. The THP1 cells are HLA-A*02:01, HLA-B*15:11, HLA-C*03:03, HLA-DRB1*01:01, HLA-DRB1*15:01, HLA-DRB5*01:01. Pure MHC will pulse THP-1 macrophages with an antigen (e.g. therapeutic biologic, subunit vaccine, small molecule drug) or a complex mixture of antigens (e.g. inactivated vaccine, bacterial lysate) in culture. The THP-1 macrophages will process and present the pulsed antigen in HLA Class I (cross-presentation) and HLA Class II. Pure MHC will purify the peptide-HLA complexes, isolate the peptide ligands, and either identify novel HLA class I or class II peptide ligands de novo and/or validate existing HLA class I/II peptides using high sensitivity LCMS.

PRM/MRM HLA Peptide Target Validation

Pure MHC will develop a custom targeted multiple reaction monitoring (MRM) liquid chromatography-mass spectrometry (LCMS) method that can accommodate up to 50 distinct peptide target sequences for detection and quantitation in various biological samples. Our approach begins with the creation of a standard curve using synthetic peptides corresponding to the targets, which enables absolute molar quantification. Next, peptide-HLA complexes are purified from the biological specimens, followed by the isolation of the peptide ligands. We then spike in synthetic peptides that match the targeted peptides and are tagged with a stable heavy isotope, to the extracted immunopeptidome. The heavy synthetics act as both a positive control and an internal normalizer. By comparing the LCMS intensity of the native peptides in the sample to that of the synthetic standard curve, we can accurately determine the molarity of the native peptides. This quantitative data provides a reliable means to estimate the abundance of peptide-HLA complexes present in the biological sample under investigation.