Dibasic endoprotease (also known as Kex2 protease, YSCF protease, EC 3.4.21.61) is responsible for processing the precursors of killer toxin and alpha-factor in yeast. It is a calcium ion-dependent serine protease that specifically recognizes and cleaves peptide bonds at the carboxyl terminus of dibasic amino acid pairs such as Arg-Arg and Lys-Arg.
This kit applies the double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) principle for the detection of recombinant dibasic endoprotease residue. The recombinant dibasic endoprotease standard (20384-B) and test samples are added to the pre-coated anti-recombinant dibasic endoprotease antibody plate (20384-A), followed by the addition of diluted biotin-labeled recombinant dibasic endoprotease detection antibody (20384-C), and finally Streptavidin-HRP (SA-HRP) (20384-D), forming an antibody + antigen + antibody-Biotin + SA-HRP complex. After plate washing, TMB chromogenic solution (20384-H) is added for color development. TMB is catalyzed by HRP enzyme from colorless to blue and finally converted to yellow under the action of stop solution (20384-I). The intensity of yellow color is positively correlated with the content of recombinant dibasic endoprotease detected in the sample. This kit can be used to determine trace residues of recombinant dibasic endoprotease in samples from recombinant protein drug or fusion expression processes using recombinant dibasic endoprotease for enzymatic cleavage.

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The performance of this kit has been fully evaluated, with a standard curve range of 1 ng/mL to 64 ng/mL, intra-assay precision <10%, and inter-assay precision <15%.