Development of a high-throughput automated analyzer using biochip array technology.
BACKGROUND
Use of protein array technology over conventional assay methods has advantages that include simultaneous detection of multiple analytes, reduction in sample and reagent volumes, and high output of test results. The susceptibility of ligands to denaturation, however, has impeded production of a stable, reproducible biochip platform, limiting most array assays to manual or, at most, semiautomated processing techniques. Such limitations may be overcome by novel biochip fabrication procedures.


METHODS
After selection of a suitable biochip substrate, biochip surfaces were chemically modified and assessed to enable optimization of biochip fabrication procedures for different test panels. The assay procedure was then automated on a dedicated instrument, and assay performance was determined for a panel of cytokine markers. Assay results were then compared with a commercial method for measurement of cytokine markers.


RESULTS
Secondary ion mass spectrometry and x-ray photoelectron spectroscopy demonstrated appropriate and reproducible modification of the biochip surface. Contact-angle studies also confirmed generation of hydrophobic surfaces that enabled containment of droplets for fabrication of discrete test regions. Automation of the biochip assays on a dedicated instrument produced excellent cytokine marker performance with intra- and interassay imprecision <10% for most analytes. Comparison studies showed good agreement with other methods (r = 0.95-0.99) for cytokines.


CONCLUSIONS
Performance data from this automated biochip array analyzer provide evidence that it is now possible to produce stable and reproducible biochips for output of more than 2000 test results per hour.
Fitzgerald
Stephen P
SP
Randox Laboratories, Crumlin, United Kingdom.
Lamont
John V
JV
McConnell
Robert I
RI
Benchikh
El O
el O
D001322:Autoanalysis; D015415:Biomarkers / Q000032:analysis; D016207:Cytokines / Q000032:analysis; D006801:Humans; D008163:Luminescent Measurements; D040081:Protein Array Analysis / Q000379:methods*; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D013052:Spectrometry, X-Ray Emission
D015415:Biomarkers; D016207:Cytokines