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- W99005642 abstract "This chapter focuses on some of the biological and practical issues related to the use of peptide/major histocompatibility complex (pMHC) tetramers. The basic components of a pMHC complex (class I) include the peptide, MHC class I molecule, and beta-2-microglobulin. For subsequent tetramerization, the MHC class I molecule is engineered with a C-terminal substrate peptide for BirA-mediated biotinylation, biotin signal peptide (bsp). This MHC l-bsp recombinant protein is generally produced using E. coli in inclusion bodies. There are now numerous human and mouse MHC I alleles engineered with bsp in efficient E. coli expression vectors. The first step when a new tetramer is made is to validate and titrate the reagent. Tetramers may fail for a number of reasons—peptides of very low MHC binding affinity do not fold into pMHC complexes well, incomplete purification or biotinylation of pMHC complexes, or bad batches of streptavidin–fluorophore . A T cell line or clone specific for the pMHC complex in the tetramer is needed for this process. The chapter discusses some of the advanced uses of pMHC tetramers. The use of MHC class I tetramers in humans is briefly described." @default.
- W99005642 created "2016-06-24" @default.
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- W99005642 date "2005-01-01" @default.
- W99005642 modified "2023-09-26" @default.
- W99005642 title "Tetramer Analysis" @default.
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- W99005642 doi "https://doi.org/10.1016/b978-012455900-4/50284-1" @default.
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