| A standard testing system of serumβ2–microglobulin(β2–m) was found by the method of serumβ2–microglobulin monoclonal antibody enzyme-linked immunosorbent assay(ELISA). Firstly,β2-m monoclonal antibody ascites was purified from the ascites protein, the methods was to acquireβ2-m monoclonal antibody after ascites protein had been purified by using method of saturated ammonium sulfate sedimentation combined with molecule griddle. Secondly,β2-microglobulin monoclonal antibody was signed by HRP: the glycoside base was oxidized to aldehyde group by sodium iodinate and reacted with amino-group joined in theβ2-microglobulin monoclonal antibody, so that the HRP recombined with theβ2-microglobulin monoclonal antibody as enzyme signed secondary antibody. At last, the protein coating concentration and enzyme labeling antibody concentration were determined through square matrix method, the optimal delution concentrations of the first antibody and the enzyme signed secondary antibody were 1:128000 and 1:1000 respectively. The cut-off value ofβ2-microglobulin monoclonal antibody ELISA , which was determined by the ROC method,was 0.35~0.4. After comparing this method with another one, the statistics analysis showed that no significant differences existed( N>40,χ2=0.551, df=1, P>0.05), so this standard ELISA detecting system have been established, which is characterized with high sensitivity, specificity and stability.
|
PDF Full Text Request