Protein analysis

摘要

A method of forming an array of proteins selected from antigens or antibodies; said method comprising the steps of (i) expressing in a recombinant cell, a fusion protein which comprises either (a) an antigen or (b) an antibody binding protein, fused to a peptide having up to 50 amino acids, which peptide comprises amino acid sequence of SEQ ID NO 1 LX₁X₂IX₃X₄X₅X₆KX₇X₈X₉X₁₀ (SEQ ID NO 1) where X₁ is a naturally occurring amino acid, X₂ is any naturally occurring amino acid other than leucine, valine, isoleucine, tryptophan, phenylalanine or tyrosine, X₃ is phenylalanine or leucine, X₄ is glutamine or asparagine, X₅ is alanine, glycine, serine or threonine, X₆ is glycine or methionine, X₇ is isoleucine, methionine or valine, X₈ is glutamine, leucine, valine, tyrosine or isoleucine, X₉ is tryptophan, tyrosine, valine, phenylalanine, leucine or isoleucine and X₁₀ is any naturally occurring amino acid other than asparagine or glutamine; where said peptide is capable of being biotinylated by a biotin ligase at the lysine residue adjacent to X₆; (ii) biotinylating said peptide of the fusion protein at the lysine residue adjacent X₆; (iii) isolating the biotinylated fusion protein; (iv) applying the biotinylated fusion protein to an avidin or streptavidin coated non-porous support; (v) forming an array of at least three different proteins on the support by either (a) where the fusion protein comprises an antigen, carrying out steps (i) to (iv) the desired number of times to form an antigen array; or (b) where the fusion protein comprises an antibody binding protein, applying to said protein, either prior to or after step (iv) a plurality of different antibodies or binding fragments thereof.