{"id":72180,"date":"2026-08-04T12:35:22","date_gmt":"2026-08-04T10:35:22","guid":{"rendered":"https:\/\/inmuno.es\/index.php\/2026\/08\/04\/peptide-exchange-competent-class-i-mhc-molecules-produced-in-eukaryotic-cells-for-rapid-production-of-mhc-multimers\/"},"modified":"2026-08-04T12:35:22","modified_gmt":"2026-08-04T10:35:22","slug":"peptide-exchange-competent-class-i-mhc-molecules-produced-in-eukaryotic-cells-for-rapid-production-of-mhc-multimers","status":"publish","type":"post","link":"https:\/\/inmuno.es\/index.php\/2026\/08\/04\/peptide-exchange-competent-class-i-mhc-molecules-produced-in-eukaryotic-cells-for-rapid-production-of-mhc-multimers\/","title":{"rendered":"Peptide exchange-competent class I MHC molecules produced in eukaryotic cells for rapid production of MHC multimers"},"content":{"rendered":"<div>\n<p><b>J Immunol<\/b>. 2026 Aug 4;215(8):vkag210. doi: 10.1093\/jimmun\/vkag210.<\/p>\n<p><b>ABSTRACT<\/b><\/p>\n<p>Class I MHC peptide (MHC-Ip) multimers are well-established reagents that detect antigen-specific T cells. The classical method for production of MHC-Ip multimers begins with the expression of MHC heavy chains (HCs) and \u03b22-microglobulin (\u03b22m) subunits as inclusion bodies in Escherichia coli and is followed by denaturant solubilization, in vitro folding in the presence of a defined peptide ligand, and purification by size exclusion chromatography. This protocol is labor intensive, difficult to scale, and represents a significant bottleneck in application of the technology. Herein, we present a novel method for the expression in eukaryotic cells of secreted peptide exchange-competent class I MHC proteins in their native conformation. In this method, expression constructs are engineered as bimolecular complexes composed of an MHC HC and a \u03b22m molecule covalently linked at its amino terminus to an MHC-binding peptide through a flexible peptide linker containing a defined protease site. Upon proteolysis, the original peptide occupant of the MHC binding site dissociates and is easily replaced with a synthetic peptide. When leucine zippers are added to the carboxyl terminus of each subunit, protease cleavage of the linker results in a stable HC\/\u03b22m complex that can be isolated and stored for subsequent peptide loading. Using this method, we have produced homogeneous MHC-Ip complexes for 25 class I MHC alleles and demonstrated that tetramers produced in this way are equivalent to conventionally produced tetramers for T-cell staining.<\/p>\n<p>PMID:<a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/42548059\/?utm_source=SimplePie&amp;utm_medium=rss&amp;utm_content=2985117R&amp;ff=20260804063522&amp;v=2.20.0.post5+40e1b98\">42548059<\/a> | DOI:<a href=\"https:\/\/doi.org\/10.1093\/jimmun\/vkag210\">10.1093\/jimmun\/vkag210<\/a><\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p>J Immunol. 2026 Aug 4;215(8):vkag210. doi: 10.1093\/jimmun\/vkag210. ABSTRACT Class I MHC peptide (MHC-Ip) multimers are well-established reagents that detect antigen-specific T cells. The classical method for production of MHC-Ip multimers begins with the expression of MHC heavy chains (HCs) and \u03b22-microglobulin (\u03b22m) subunits as inclusion bodies in Escherichia coli and is followed by denaturant solubilization, &#8230; <a title=\"Peptide exchange-competent class I MHC molecules produced in eukaryotic cells for rapid production of MHC multimers\" class=\"read-more\" href=\"https:\/\/inmuno.es\/index.php\/2026\/08\/04\/peptide-exchange-competent-class-i-mhc-molecules-produced-in-eukaryotic-cells-for-rapid-production-of-mhc-multimers\/\" aria-label=\"Read more about Peptide exchange-competent class I MHC molecules produced in eukaryotic cells for rapid production of MHC multimers\">Read more<\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[42,71],"tags":[],"class_list":["post-72180","post","type-post","status-publish","format-standard","hentry","category-publicaciones","category-the-journal-of-immunology"],"_links":{"self":[{"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/posts\/72180","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/comments?post=72180"}],"version-history":[{"count":0,"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/posts\/72180\/revisions"}],"wp:attachment":[{"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/media?parent=72180"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/categories?post=72180"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/inmuno.es\/index.php\/wp-json\/wp\/v2\/tags?post=72180"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}