An enzyme immunoassay for the quantitative measurement of NOV in serum, plasma, and urine.
The CCN family of genes constitutes six members of small secreted cysteine-rich proteins, which exists only in vertebrates. The major members of CCN are CCN1 (Cyr61), CCN2 (CTGF), and CCN3 (NOV). CCN4, CCN5, and CCN6 were formerly reported to be in the Wisp family, but they are now integrated into CCN family due to the resemblance of their four principal modules: insulin like growth factor binding protein, von Willebrand factor typeC, thrombospondin type 1, and carboxy-terminal cysteine knot domain (1). The CCN proteins have been shown to have key roles as matricellular proteins, serving as adaptor molecules connecting the cell surface and extracellular matrix (ECM). CCN proteins not only regulate crucial biological processes including cell differentiation, proliferation, adhesion, migration, apoptosis, ECM production, chondrogenesis and angiogenesis, but also have more sinister roles promoting conditions such as fibrogenesis (2). Altered NOV expression has been reported for glioma cells (3). In addition, NOV mRNA and protein expression was shown to be lower in childhood adrenocortical tumors than in normal adrenal cortex while no significant difference was observed between adenomas and carcinomas (4,5). Furthermore, low levels of NOV were also seen in breast cancer patients with poor prognosis and mortality and with significantly decreased survival (6).
The DRG NOV ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA) based on the sandwich principle. The microtiter wells are coated with a monoclonal
[mouse] antibody directed towards a unique antigenic site of the NOV molecule.An aliquot of patient sample containing endogenous NOV is incubated in the coated well together with assay buffer. After washing, a biotinylated anti-NOV antibody (enzyme conjugate) is added. After incubation, the unbound conjugate is washed off, and Streptavidin conjugated with horseradish peroxidase (Enzyme Complex) is added. After incubation, unbound enzyme complex is washed off. The amount of bound peroxidase is proportional to the concentration of NOV in the sample. Having added the substrate solution, the intensity of colour developed is proportional to the concentration of NOV in the patient sample.