Construction and evaluation of an automated light directed protein-detecting microarray synthesizer.
We have designed, constructed, and evaluated an automated instrument that has produced high-density arrays with more than 30 000 peptide features within a 1.5 cm(2) area of a glass slide surface. These arrays can be used for high throughput library screening for protein binding ligands, for potential drug candidate molecules, or for discovering biomarkers. The device consists of a novel fluidics system, a relay control electrical system, an optics system that implements Texas Instruments' digital micromirror device (DMD), and a microwave source for accelerated synthesis of peptide arrays. The instrument implements two novel solid phase chemical synthesis strategies for producing peptide and peptoid arrays. Biotin-streptavidin and DNP anti-DNP (dinitrophenol) models of antibody small molecule interactions were used to demonstrate and evaluate the instrument's capability to produce high-density protein detecting arrays. Several screening assay and detection schemes were explored with various levels of efficiency and assays with sensitivity of 10 nM were also possible.
Marthandan
N
N
McDermott Center for Human Growth and Development, University of Texas, South Western Medical Center, Dallas, TX 75390-8573, USA. nishanth.mm@gmail.com
Klyza
S
S
Li
S
S
Kwon
Y U
YU
Kodadek
T
T
Garner
H R
HR
D004867:Equipment Design; D019544:Equipment Failure Analysis; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008872:Microwaves; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods; D055095:Optics and Photonics / Q000295:instrumentation; D010783:Photometry / Q000295:instrumentation* / Q000379:methods; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012371:Robotics / Q000295:instrumentation* / Q000379:methods; D012680:Sensitivity and Specificity