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For this reason, we have developed a method using homogeneously loaded peptide\u2013MHC class II complexes to generate and select specific mAb reactive with these structures using hen egg lysozyme (HEL) and I-A<jats:sup>k<\/jats:sup>as a model system. mAbs specific for either HEL-(46\u201361)\u2013A<jats:sup>k<\/jats:sup>or HEL-(116\u2013129)\u2013A<jats:sup>k<\/jats:sup>have been isolated. They cross-react with a small subset of I-A<jats:sup>k<\/jats:sup>molecules loaded with self peptides but can nonetheless be used for flow cytometry, immunoprecipitation, Western blotting, and intracellular immunofluorescence to detect specific HEL peptide\u2013MHC class II complexes formed by either peptide exposure or natural processing of native HEL. An example of the utility of these reagents is provided herein by using one of the anti-HEL-(46\u201361)\u2013A<jats:sup>k<\/jats:sup>specific mAbs to visualize intracellular compartments where I-A<jats:sup>k<\/jats:sup>is loaded with HEL-derived peptides early after antigen administration. Other uses, especially for<jats:italic>in vivo<\/jats:italic>tracking of specific ligand-bearing antigen-presenting cells, are discussed.<\/jats:p>","DOI":"10.1073\/pnas.94.25.13856","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T14:40:52Z","timestamp":1027694452000},"page":"13856-13861","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":113,"title":["Production, specificity, and functionality of monoclonal antibodies to specific peptide\u2013major histocompatibility complex class II complexes formed by processing of\u2009exogenous\u2009protein"],"prefix":"10.1073","volume":"94","author":[{"given":"Guangming","family":"Zhong","sequence":"first","affiliation":[{"name":"Lymphocyte Biology Section, Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 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