Final Report Summary - PROCHEM (Protein stabilization by chemistry: total synthesis of an MHC class I scaffold)
Summary report:
Cancer immunotherapy benefits from technology for T cell visualization and T cell depletion/enrichment. Techniques that currently allow visualization of antigen-specific T cell responses involve biotinylation of soluble MHC monomers loaded with a peptide of choice and subsequent tetramerization by binding to a streptavidin. Currently, availability and reliability of the peptide-Major Histocompatibility Complex (pMHC) reagents dictates their development and subsequent applications. Recently, the development of MHC tetramers with exchangeable peptide ligand has partly solved the problem of the tedious and laborious preparation of individual MHCs. However, the development of an improved technology suitable for high throughput screening and ultimately of empty stable MHCs is highly desirable. PROCHEM’s main goal is to design and prepare, by chemical total synthesis, a truncated version of the MHC class 1 protein based on the human allele HLA::A2.1 that aims to furnish a practical tool to detect antigen specific T cells. Ultimately, we want to generate a stable empty MHC (α1,α2) platform by using chemical modification along the peptide sequence and by circularization of the platform. We use solid phase peptide synthesis to prepare peptide fragments that, after being properly functionalized, are connected to each other by mean of native chemical ligation and click reaction to furnish the desired MHC (α1,α2) platform. The sequence of the synthesized platform has been confirmed by SDS PAGE analysis and MSMS.
Cancer immunotherapy benefits from technology for T cell visualization and T cell depletion/enrichment. Techniques that currently allow visualization of antigen-specific T cell responses involve biotinylation of soluble MHC monomers loaded with a peptide of choice and subsequent tetramerization by binding to a streptavidin. Currently, availability and reliability of the peptide-Major Histocompatibility Complex (pMHC) reagents dictates their development and subsequent applications. Recently, the development of MHC tetramers with exchangeable peptide ligand has partly solved the problem of the tedious and laborious preparation of individual MHCs. However, the development of an improved technology suitable for high throughput screening and ultimately of empty stable MHCs is highly desirable. PROCHEM’s main goal is to design and prepare, by chemical total synthesis, a truncated version of the MHC class 1 protein based on the human allele HLA::A2.1 that aims to furnish a practical tool to detect antigen specific T cells. Ultimately, we want to generate a stable empty MHC (α1,α2) platform by using chemical modification along the peptide sequence and by circularization of the platform. We use solid phase peptide synthesis to prepare peptide fragments that, after being properly functionalized, are connected to each other by mean of native chemical ligation and click reaction to furnish the desired MHC (α1,α2) platform. The sequence of the synthesized platform has been confirmed by SDS PAGE analysis and MSMS.