Direct detection of antibody-antigen binding using an on-chip artificial pore.
We demonstrate a rapid and highly sensitive all-electronic technique based on the resistive pulse method of particle sizing with a pore to detect the binding of unlabeled antibodies to the surface of latex colloids. Here, we use an on-chip pore to sense colloids derivatized with streptavidin and measure accurately their diameter increase on specific binding to several different types of antibodies. We show the sensitivity of this technique to the concentration of free antibody and that it can be used to perform immunoassays in both inhibition and sandwich configurations. Overall, our technique does not require labeling of the reactants and is performed rapidly by using very little solution, and the pore itself is fabricated quickly and inexpensively by using soft lithography. Finally, because this method relies only on the volume of bound ligand, it can be generally applied to detecting a wide range of ligand-receptor binding reactions.
Saleh
Omar A
OA
Department of Physics, Princeton University, Princeton, NJ 08544, USA.
Sohn
Lydia L
LL
D000937:Antigen-Antibody Reactions; D003102:Colloids / Q000737:chemistry; D004306:Dose-Response Relationship, Immunologic; D004581:Electronics; D007118:Immunoassay; D007840:Latex / Q000737:chemistry; D008024:Ligands; D008962:Models, Theoretical; D011485:Protein Binding; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000737:chemistry; D013997:Time Factors
D003102:Colloids; D007840:Latex; D008024:Ligands; D019809:Streptavidin