Improved continuous-flow print head for micro-array deposition.
Limitations in depositing ligands using conventional micro-array pin spotting have hindered the application of surface plasmon resonance imaging (SPRi) technology. To address these challenges we introduce a modification to our continuous-flow micro-spotting technology that improves ligand deposition. Using Flexchip protein A/G and neutravidin capturing surfaces, we demonstrate that our new microfluidic spotter requires 1000 times less concentrated antibodies and biotinylated ligands than is required for pin spotting. By varying the deposition flow rate, we show that the design of our tip overlay flow cell is efficient at delivering sample to the substrate surface. Finally, contact time studies show that it is possible to capture antibodies and biotinylated ligands at concentrations of less than 0.1 ug/ml and 100 pM, respectively. These improvements in spotting technology will help to expand the applications of SPRi systems in areas such as antibody screening, carbohydrate arrays, and biomarker detection.
Eddings
Mark A
MA
Department of Bioengineering, University of Utah, Salt Lake City, UT 84132, USA.
Miles
Adam R
AR
Eckman
Josh W
JW
Kim
Jungkyu
J
Rich
Rebecca L
RL
Gale
Bruce K
BK
Myszka
David G
DG
D053000:Analytic Sample Preparation Methods; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000276:immunology; D001710:Biotin / Q000378:metabolism; D050260:Carbohydrate Metabolism; D002417:Cattle; D008024:Ligands; D040081:Protein Array Analysis / Q000379:methods*; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance / Q000379:methods*; D013997:Time Factors
D000906:Antibodies; D000941:Antigens; D008024:Ligands; D001710:Biotin