Expression and characterization of soluble epitope-defined major histocompatibility complex (MHC) from stable eukaryotic cell lines
In: Journal of Immunological Methods, Jg. 464 (2019), S. 22-30
Online
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Zugriff:
MHC class I-specific reagents such as fluorescently-labeled multimers (e.g., tetramers) have greatly advanced the understanding of CD8+ T cells under normal and diseased states. However, recombinant MHC class I components (comprising MHC class I heavy chain and β2 microglobulin) are usually produced in bacteria following a lengthy purification protocol that requires additional non-covalent folding steps with exogenous peptide for complete molecular assembly. We have provided an alternative and rapid approach to generating soluble and fully-folded MHC class I molecules in eukaryotic cell lines (such as CHO cells) using a Sleeping Beauty transposon system. Importantly, this method culminates in generating stable cell lines that reliably secrete epitope-defined MHC class I molecules into the tissue media for convenient purification and eventual biotinylation/multimerization. Additionally, MHC class I components are covalently linked, providing the opportunity to produce a diverse set of CD8+ T cell-specific reagents bearing peptides with various affinities to MHC class I.
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Expression and characterization of soluble epitope-defined major histocompatibility complex (MHC) from stable eukaryotic cell lines
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Autor/in / Beteiligte Person: | Anderson, Trevor S. ; Wooster, Amanda L. ; Lowe, Devin B. |
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Zeitschrift: | Journal of Immunological Methods, Jg. 464 (2019), S. 22-30 |
Veröffentlichung: | Elsevier BV, 2019 |
Medientyp: | unknown |
ISSN: | 0022-1759 (print) |
DOI: | 10.1016/j.jim.2018.10.006 |
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