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Time-resolved Chemiluminescence Technique for the Microdetermination of Proteins Based on Their Complexation with Copper(II)

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Abstract

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 µg/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.

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Authors and Affiliations

  1. College of Chemistry and Environment Science, Hebei University, Baoding, 071002, Hebei Province, P. R. China

    Zheng Ping Li & Li Na Zhang

  2. Department of Chemistry, Chengde Teacher’s College for Nationalities, Chengde, 067000, Hebei Province, P. R. China

    Shuang Shuang Fan

  3. Hebei Entry and Exit Inspection and Quarantine Bureau of P. R. China, Shijiazhuang, 051000, Hebei Province, P. R. China

    Feng Chi Wang

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  1. Zheng Ping Li
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  2. Shuang Shuang Fan
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  3. Li Na Zhang
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  4. Feng Chi Wang
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Correspondence to Zheng Ping Li.

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Li, Z.P., Fan, S.S., Zhang, L.N. et al. Time-resolved Chemiluminescence Technique for the Microdetermination of Proteins Based on Their Complexation with Copper(II). ANAL. SCI. 20, 1327–1331 (2004). https://doi.org/10.2116/analsci.20.1327

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  • DOI: https://doi.org/10.2116/analsci.20.1327

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