Development of Miniaturized Competitive Immunoassays on a Protein Chip as a Screening Tool for Drugs

Abstract

Background: Doping in sports has become a serious problem. Gas chromatography-mass spectrometry (GC-MS) serves as an effective reference method, but it is limited by low throughput and is therefore not suitable for large-scale screening. Use of protein chips for highthroughput screening of all athletes for prohibited substances could become an important complementary tool to GC-MS. Methods: We developed a protein chip based on an aldehyde-activated glass slide containing 10 physically isolated arrays. The chip was used to screen urine from 1347 athletes for prohibited substances and to screen a negative control group consisting of 200 females and 120 males. Urine samples from 66 individuals known to be abusers, provided by the China Doping Control Center (CDCC), and 129 standard prohibited substances were tested as positive controls. Results: All 1347 urine samples screened by means of the protein chips were also subjected to reference analysis by GC-MS at the CDCC. There was no qualitative difference between the results obtained with the two methods. The correlation coefficient (r 2 ) for the quantitative results obtained with the protein chip and GC-MS was 0.991.

Protein chips for high-throughput doping screening in athletes

“…The results of the chip were confirmed by the reference analysis (GC/MS) at the China Doping Control Center (one of the WADA-accredited laboratories) in Beijing. The correlation coefficient (r 2 ) for the quantitative results obtained with the protein chip and GC/MS was 0.991 [18].…”

“…The assay was repeated in a 96-well plate and it was found that all the exogenous analytes, including amphetamine, had a low PBS/blank urine ratio (1.13-1. 18). This suggests that some nondoping substances in human urine may have interfered with the interaction of amphetamine and its antibody [18].…”

“…18). This suggests that some nondoping substances in human urine may have interfered with the interaction of amphetamine and its antibody [18].…”

Up-to-Date Applications of Microarrays and Their Way to Commercialization

“…Du et al [31,32,36] developed a Protein Microarray for the detection of different prohibited drugs in various biological fluids. They printed up to ten different drug conjugates in triplicates onto an aldehyde modified glass slide including mouse IgG as internal control.…”

Construction of a multiplex allele-specific PCR-based universal array (ASPUA) and its application to hearing loss screening

“…In the recent few years microarray technology has had many applications in the areas of disease diagnosis and drug discovery. For example, we have developed protein microarrays for small molecule screening and severe acute respiratory syndrome (SARS) patient sample analysis [Du et al, 2005;Kong et al, 2005], and also DNA microarrays for the early detection of SARS coronavirus as well as for the potential side effect study in developing siRNAbased hepatitis B virus (HBV) inhibiting drugs Guo et al, 2005]. DNA microarrays permit high-throughput mutation detection approaches, such as allele-specific primer extension on microarrays [Pastinen et al, 2000], PCR/ligase detection reaction (LDR) universal array [Gerry et al, 1999], microsphere-based single-base chain extension (SBCE) [Chen et al, 2000], sequence-tagged molecular inversion probes (MIP) [Hardenbol et al, 2003], and combinatorial sequencing-byhybridization (cSBH) [Cowie et al, 2004].…”