Methods for producing functionalized immunoglobulin superfamily proteins

摘要

The present invention relates to a process of producing a functional immunoglobulin superfamily protein, which has at least one disulphide bond when functional, the process comprising the steps of providing a bacterial cell comprising a gene coding for the protein, the gene is expressible in said cell, cultivating the cell under conditions where the gene is expressed, isolating the protein from the cell without reducing it, and subjecting the isolated protein to a folding treatment. Preferably, the immunoglobulin superfamily protein is selected from the group consisting of antibodies, immunoglobulin variable (V) regions, immunoglobulin constant (C) regions, immunoglobulin light chains, immunoglobulin heavy chains, CD1, CD2, CD3, Class I and Class II histocompability molecules, beta 2microglobulin ( beta 2m), lymphocyte function associated antigen-3 (LFA-3) and Fc gamma RIII, CD7, CD8, Thy-1 and Tp44 (CD28), T cell receptor, CD4, polyimmunoglobulin receptor, neuronal cell adhesion molecule (NCAM), myelin associated glycoprotein (MAG), P myelin protein, carcino-embryonic antigen (CEA), platelet derived growth factor receptor (PDGFR), colony stimulating factor-1 receptor, alpha beta -glycoprotein, ICAM (intercellular adhesion molecule), platelet and interleukins. Important embodiments of the invention is a stable peptide free MHC protein obtainable by a process of the invention and a kit comprising a MHC class I heavy chain and a beta 2m allowing the recipient to produce and measure or detect a functional MHC class I protein to which a peptide, which is capable of binding to said MHC class I protein, can be added leading to the generation of a functional MHC class I protein.