Method
The invention provides a method of detecting the presence of anti-MHC antibodies in a sample comprising contacting said sample with one or more recombinant MHC molecules or functionally equivalent variants, derivatives or fragments thereof and detecting the binding or absence of binding of antibodies to said recombinant MHC molecules. This method allows the detection and/or identification of one or more specific MHC particularly HLA allele antibodies
1 . A method of detecting the presence of one or more allele specific anti-Major Histo-Compatibility Complex (MHC) antibodies in a body fluid sample comprising contacting said sample with one or more recombinant MHC molecules which each bind to a different allele specific MHC antibody, if present in said sample, and detecting binding or absence of binding of said one or more allele specific antibodies to said recombinant MHC molecules, wherein each of said one or more allele specific antibodies is specific for a particular naturally occurring MHC allele and binds to only one of said one or more recombinant MHC molecules which contains one or more epitopes of said naturally occurring MHC allele.
2 . A method of detecting the presence of one or more allele specific anti-Human Leukocyte Antigen (HLA) antibodies in a body fluid sample comprising contacting said sample with one or more recombinant HLA molecules which each bind a different allele specific HLA antibody, if present in said sample, and detecting binding or absence of binding of said one or more allele specific antibodies to said recombinant HLA molecules, wherein each of said one or more specific antibodies is each specific for a particular HLA allele and binds to only one of said one or more recombinant HLA molecules which contains one or more epitopes of said naturally occurring HLA allele.
3 . The method as claimed in claim 1 or 2 wherein said MHC or HLA molecule is a Class I MHC or HLA molecule.
4 . The method as claimed in claim 3 , wherein said recombinant MHC or HLA molecule comprises a heavy chain and wherein the heavy chain of said MHC or HLA molecule is recombinant.
5 . The method as claimed in claim 1 or 2 wherein the antibodies which are detected are IgG, IgM or IgA.
6 . The method as claimed in claim 3 wherein said recombinant MHC or HLA molecule comprises a heavy chain, β 2 -microglobulin and peptide.
7 . A method as claimed in claim 6 wherein said peptide is derived from HIV, HCV or an influenza virus.
8 . The method of claim 1 wherein the MHC molecules are attached to a solid support.
9 . The method of claim 2 wherein the HLA molecules are attached to a solid support.
10 . The method of claim 9 or 10 wherein said solid support is a spherical bead.
11 . The method of claim 9 or 10 wherein said solid support is a nitrocellulose strip.
12 . The method of claim 9 or 10 wherein said solid support is an ELISA plate.
13 . The method of claim 1 wherein the recombinant MHC is synthesized in a prokaryotic expression system.
14 . The method of claim 2 wherein the recombinant HLA is synthesized in a prokaryotic expression system.
15 . The method of claim 1 or claim 2 wherein the body fluid sample is blood or a blood-derived sample.
16 . The method as defined in claim 1 or claim 2 wherein the bound antibody is detected by an immunosorbent assay using an antibody conjugated to a signaling means.
17 . A kit comprising at least the following components:
(a) one or more recombinant MHC molecules which each bind to a different allele specific MHC antibody, wherein each of said one or more allele specific antibodies is specific for a particular naturally occurring MHC allele and binds to only one of said one or more recombinant MHC molecules which contains one or more epitopes of said naturally occurring MHC allele; and
(b) a means for detecting anti-MHC antibodies.
18 . A kit as claimed in claim 17 which further comprises a solid support together with means for attachment of the MHC molecules.
19 . A kit as claimed in claim 17 wherein the means for detecting said anti-MHC antibodies comprises an antibody which binds to the complex formed between said MHC molecules and naturally occurring antibodies to said molecules.
20 . A kit as claimed in claim 18 , wherein said solid support is a spherical bead.
21 . A method as claimed in claim 1 or 2 wherein said recombinant MHC or HLA molecule is in the form of a fusion protein.
22 . A method as claimed in claim 21 , wherein said recombinant MHC or HLA molecule is fused to a means of immobilization.
23 . A method as claimed in claim 22 , wherein said means of immobilization is biotin.