Quantitative label-free screening for antibodies using scattering biophotonic microarray imaging.
A biophotonic array based on gold nanoparticles functionalized with antigen proteins has been used to determine the concentrations of the respective antibodies in solution. Four proteins-fibrinogen, bovine serum albumin, transferrin, and C-reactive protein-were used to construct a test array with the assay repeated a number of times. The antibody-antigen association and dissociation rate constants were determined for the antibody assays from a series of calibration experiments. The label-free determination of the unknown antibody concentrations was performed using two related kinetic analyses. From these results, the current array assay sensitivity is 250 ng ml(-1) with an accuracy of 15% using an 8-min kinetic measurement and a 16-spot averaged assay.
Olkhov
Rouslan V
RV
School of Biosciences, University of Exeter, Exeter EX4 4QD, UK.
Shaw
Andrew M
AM
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D000941:Antigens / Q000276:immunology; D015374:Biosensing Techniques; D002417:Cattle; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007700:Kinetics; D046228:Microarray Analysis / Q000379:methods*; D017785:Photons; D013194:Staining and Labeling
D000906:Antibodies; D000941:Antigens