title: Perfusion chromatography for very rapid purification of class I and II MHC proteins.
abstract: Major histocompatibility complex (MHC) proteins are surface glycoproteins that are strongly associated with either self or foreign peptides. Their interaction with the T-cell receptor on the T-cells initiates an immune response and help in discriminating between self and non-self, respectively. We describe here a novel means of rapidly purifying human MHC molecules on either small scale or large scale from the cell lysate of lymphoblastoid B cell line and from insect cell culture supernatants by using affinity perfusion chromatography. As representative cases HLA-B2705, a class I MHC molecule, and HLA-DR1, a class II MHC molecule were purified from EBV-transformed human lymphoblastoid B cells, LG2. Soluble HLA-DR1 was also purified from the cell culture supernatant of insect cells. The peptides eluted from the purified HLA-B2705 were pool sequenced and found to have the same motif as has previously been published. This new method provides a very rapid means of purifying MHC protein molecules, applicable to both large scale and small scale purification, which in turn greatly enhances the accuracy of further analysis of the associated peptides through mass spectrometry.
authors.affiliation: Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
authors.forename: P
authors.initials: P
authors.lastname: Malik
authors.forename: J L
authors.initials: JL
authors.lastname: Strominger
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D002846:Chromatography, Affinity / Q000379:methods*; D015796:HLA-B27 Antigen / Q000276:immunology / Q000302:isolation & purification*; D015798:HLA-DR1 Antigen / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D051379:Mice; D013997:Time Factors
chemical_list: D015796:HLA-B27 Antigen; D015798:HLA-DR1 Antigen