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One approach to the analysis of leucocyte cell surface proteins is to express their domains with part of another protein as a carrier. We report the use of two immunoglobulin superfamily (IgSF) domains from rat CD4 (CD4d3 + 4) in producing domains from various superfamilies as chimeric proteins in Chinese hamster ovary cell lines. Four types of construct were successfully expressed containing: (i) the two IgSF domains of CD48; (ii) the IgSF domain of mb-1 which is part of the B cell antigen recognition complex; (iii) a T cell receptor V domain; and (iv) the N-terminal domain of CD5 which belongs to the scavenger receptor superfamily. This CD5 chimeric protein was antigenic for a panel of CD5 mAbs showing that mAbs with functional effects reacted with the N-terminal domain of CD5. The CD48 chimeric protein has been used both as multivalent complexes produced by cross-linking with mAbs recognizing CD4 and in a monomeric form to analyse the kinetics of the interaction between CD48 and CD2 [van der Merwe et al. (1993) EMBO J., 12, 4945-4954].

Type

Journal article

Journal

Protein Eng

Publication Date

04/1994

Volume

7

Pages

515 - 521

Keywords

Animals, Antibodies, Monoclonal, Antibody Formation, Antigens, CD, Antigens, CD4, Antigens, CD5, Antigens, CD79, Base Sequence, CD48 Antigen, CHO Cells, Cricetinae, Genetic Vectors, Humans, Immunoglobulins, Ligands, Membrane Glycoproteins, Membrane Proteins, Molecular Sequence Data, Rats, Receptors, Antigen, B-Cell, Receptors, Antigen, T-Cell, Receptors, Immunologic, Receptors, Lipoprotein, Receptors, Scavenger, Recombinant Fusion Proteins, Scavenger Receptors, Class B