Time-resolved chemiluminescence technique for the microdetermination of proteins based on their complexation with copper(II).
Based on the complexation between proteins and Cu(II) coupled with the time-resolved chemiluminescence (CL) technique, a highly sensitive and quantitative assay for measuring proteins in solution is described. The complexes of proteins with Cu(II) have a strongly catalytic effect on the luminol-H2O2 CL reaction. Because the CL emission produced by the complexes is much more long-lived than that by Cu(II), the CL signals originating from proteins can be easily identified and measured with a time-resolved technique. On this basis, bovine albumin fraction V (BAF V) can be quantitatively determined in the range of 0.01 - 5.0 microg/ml with a detection limit of 5.8 ng/ml. The results show that the proposed assay exhibits a small variation in the response values for the same amount of different proteins, as compared to the Lowry as well as Bradford assays. The CL assay has also been studied for the detection of immobilized proteins.
College of Chemistry and Environment Science, Hebei University, Baoding 071002, Hebei Province, P R. China. lzpbd@mail.hbu.edu.cn
Zheng Ping
ZP
Li
Shuang Shuang
SS
Fan
Li Na
LN
Zhang
Feng Chi
FC
Wang
D002138:Calibration; D002384:Catalysis; D003300:Copper / Q000737:chemistry*; D007202:Indicators and Reagents; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008165:Luminol / Q000737:chemistry; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity
D007202:Indicators and Reagents; D011506:Proteins; D008165:Luminol; D003300:Copper