title: Comparative characterization of recombinant ZZ protein-alkaline phosphatase and its application in enzyme immunoassays.
abstract: A functional fusion protein, which consists of an antibody and an enzyme that can be used in enzyme immunoassays, has been constructed. However, a quantitative comparison of the characteristics of fusion proteins and chemical conjugates of the parents, which are functionally produced in a uniform microbial system, has not been adequately achieved. In this study, a fusion protein between the ZZ protein and Escherichia coli alkaline phosphatase (AP) and the parental ZZ protein and AP for chemical conjugate was functionally produced in the same bacterial system. A detailed examination of the ZZ-AP fusion protein and the effect of the ZZ-AP chemical conjugate on IgG affinity and enzymatic activity were performed. Compared with the parents, the equilibrium dissociation constant of ZZ-AP conjugate decreased by 32 % and catalytic activity decreased by 24 %, whereas the ZZ-AP fusion retained full parental activities and exhibited an approximately tenfold higher sensitivity than that of ZZ-AP conjugate in enzyme-linked immunosorbent assay. Thus, ZZ-AP fusion is a promising immunoreagent for IgG detection and a potential biolinker between antibodies and reporter enzymes (i.e., IgG-ZZ-AP fusion complex) in immunoassays.
authors.affiliation: School of Pharmacy & Bioscience, Weifang Medical University, 261053, Weifang, People's Republic of China.
authors.forename: Jin-Bao
authors.initials: JB
authors.lastname: Tang
authors.forename: Hong-Ming
authors.initials: HM
authors.lastname: Yang
authors.forename: Shu-Juan
authors.initials: SJ
authors.lastname: Liang
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Qing-Jie
authors.initials: QJ
authors.lastname: Mu
authors.forename: Jin-Bao
authors.initials: JB
authors.lastname: Zhang
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics; D000915:Antibody Affinity; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D007124:Immunoenzyme Techniques / Q000379:methods; D007074:Immunoglobulin G / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase