Clinical Chemistry
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Clinical Chemistry 52: 2273-2280, 2006. First published October 19, 2006; 10.1373/clinchem.2006.073569
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(Clinical Chemistry. 2006;52:2273-2280.)
© 2006 American Association for Clinical Chemistry, Inc.

Automation and Analytical Techniques

Rapid and Simultaneous Quantification of 4 Urinary Proteins by Piezoelectric Quartz Crystal Microbalance Immunosensor Array

Yang Luo1, Ming Chen1, Qianjun Wen2, Meng Zhao1, Bo Zhang1, Xiaoyan Li3, Feng Wang1, Qing Huang1, Chunyan Yao1, Tianlun Jiang1, Guoru Cai4 and Weiling Fu1,a

1 Department of Laboratory Medicine, 2 Laboratory Center of Urology, and 3 Endocrinology Department, Southwest Hospital, The Third Military Medical University, Chongqing, People’s Republic of China.
4 The 26th Research Institute, Chinese Electronic Scientific and Technical Group Company, Chongqing, People’s Republic of China.

aAddress correspondence to this author at: Department of Laboratory Medicine, Southwest Hospital, Third Military Medical University, Chongqing 400038, People’s Republic of China. Fax 86-23-65460909; e-mail weilingfu{at}yahoo.com.

Background: Urinary proteins are predictive and prognostic markers for diabetes nephropathy. Conventional methods for the quantification of urinary proteins, however, are time-consuming, and most require radioactive labeling. We designed a label-free piezoelectric quartz crystal microbalance (QCM) immunosensor array to simultaneously quantify 4 urinary proteins.

Methods: We constructed a 2 x 5 model piezoelectric immunosensor array fabricated with disposable quartz crystals for quantification of microalbumin, {alpha}1-microglobulin, ß2-microglobulin, and IgG in urine. We made calibration curves after immobilization of antibodies at an optimal concentration and then evaluated the performance characteristics of the immunosensor with a series of tests. In addition, we measured 124 urine samples with both QCM immunosensor array and immunonephelometry to assess the correlation between the 2 methods.

Results: With the QCM immunosensor array, we were able to quantify 4 urinary proteins within 15 min. This method had an analytical interval of 0.01–60 mg/L. The intraassay and interassay imprecisions (CVs) were <10%, and the relative recovery rates were 90.3%–109.1%. Nonspecificity of the immunosensor was insignificant (frequency shifts <20 Hz). ROC analyses indicated sensitivities were ≥95.8% and, specificities were ≥76.3%. Bland–Altman difference plots showed the immunosensor array to be highly comparable to immunonephelometry.

Conclusions: The QCM system we designed has the advantages of being rapid, label free, and highly sensitive and thus can be a useful supplement to commercial assay methods in clinical chemistry.




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A. A. Ellington, I. J. Kullo, K. R. Bailey, and G. G. Klee
Antibody-Based Protein Multiplex Platforms: Technical and Operational Challenges
Clin. Chem., February 1, 2010; 56(2): 186 - 193.
[Abstract] [Full Text] [PDF]


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