NDLI: Competitive Binding to Cuprous Ions of Protein and BCA in the Bicinchoninic Acid Protein Assay

Content Provider	PubMed Central
Author	Huang, Tao Long, Mian Huo, Bo
Copyright Year	2010
Abstract	Although Bicinchoninic acid (BCA) has been widely used to determine protein concentration, the mechanism of interaction between protein, copper ion and BCA in this assay is still not well known. Using the Micro BCA protein assay kit (Pierce Company), we measured the absorbance at 562 nm of BSA solutions with different concentrations of protein, and also varied the BCA concentration. When the concentration of protein was increased, the absorbance exhibited the known linear and nonlinear increase, and then reached an unexpected plateau followed by a gradual decrease. We introduced a model in which peptide chains competed with BCA for binding to cuprous ions. Formation of the well-known chromogenic complex of BCA-Cu1+-BCA was competed with the binding of two peptide bonds (NTPB) to cuprous ion, and there is the possibility of the existence of two new complexes. A simple equilibrium equation was established to describe the correlations between the substances in solution at equilibrium, and an empirical exponential function was introduced to describe the reduction reaction. Theoretical predictions of absorbance from the model were in good agreement with the measurements, which not only validated the competitive binding model, but also predicted a new complex of BCA-Cu1+-NTPB that might exist in the final solution. This work provides a new insight into understanding the chemical bases of the BCA protein assay and might extend the assay to higher protein concentration.
Related Links	http://dx.doi.org/10.2174/1874120701004010271
Ending Page	278
Page Count	8
Starting Page	271
File Format	PDF
ISSN	18741207
e-ISSN	18741207
Journal	The Open Biomedical Engineering Journal
Volume Number	4
Language	English
Publisher	Bentham Open
Publisher Date	2010-11-30
Access Restriction	Open
Rights Holder	Bentham Open
Subject Keyword	Medicine (miscellaneous) Bioengineering Biomedical Engineering Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Medicine Bioengineering Biomedical Engineering

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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