Decoupling competing surface binding kinetics and reconfiguration of receptor footprint for ultrasensitive stress assays.
Cantilever arrays have been used to monitor biochemical interactions and their associated stress. However, it is often necessary to passivate the underside of the cantilever to prevent unwanted ligand adsorption, and this process requires tedious optimization. Here, we show a way to immobilize membrane receptors on nanomechanical cantilevers so that they can function without passivating the underlying surface. Using equilibrium theory, we quantitatively describe the mechanical responses of vancomycin, human immunodeficiency virus type 1 antigens and coagulation factor VIII captured on the cantilever in the presence of competing stresses from the top and bottom cantilever surfaces. We show that the area per receptor molecule on the cantilever surface influences ligand-receptor binding and plays an important role on stress. Our results offer a new way to sense biomolecules and will aid in the creation of ultrasensitive biosensors.
London Centre for Nanotechnology and Departments of Medicine and Physics, University College London, 17-19 Gordon Street, London WC1H 0AH, UK.|Department of Materials, Imperial College London, London SW7 2AZ, UK.
Samadhan B
SB
Patil
London Centre for Nanotechnology and Departments of Medicine and Physics, University College London, 17-19 Gordon Street, London WC1H 0AH, UK.
Manuel
M
Vögtli
London Centre for Nanotechnology and Departments of Medicine and Physics, University College London, 17-19 Gordon Street, London WC1H 0AH, UK.|Division of Infection &Immunity, University College London, Cruciform Building, Gower Street, London WC1E 6BT, UK.
Benjamin
B
Webb
UCL Institute for Liver and Digestive Health, Royal Free Hospital, London NW3 2QG, UK.
Giuseppe
G
Mazza
UCL Institute for Liver and Digestive Health, Royal Free Hospital, London NW3 2QG, UK.
Massimo
M
Pinzani
Department of Materials, Imperial College London, London SW7 2AZ, UK.
Yeong-Ah
YA
Soh
London Centre for Nanotechnology and Departments of Medicine and Physics, University College London, 17-19 Gordon Street, London WC1H 0AH, UK.
Rachel A
RA
McKendry
London Centre for Nanotechnology and Departments of Medicine and Physics, University College London, 17-19 Gordon Street, London WC1H 0AH, UK.|Department of Chemistry, Jomo Kenyatta University of Agriculture and Technology, PO Box 62000, Nairobi, Kenya.
Joseph W
JW
Ndieyira
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism; D002161:Camelids, New World; D004867:Equipment Design; D005169:Factor VIII / Q000378:metabolism; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism*; D061905:Single-Domain Antibodies / Q000276:immunology; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*; D013499:Surface Properties; D014640:Vancomycin / Q000378:metabolism; D054299:env Gene Products, Human Immunodeficiency Virus / Q000276:immunology
D000900:Anti-Bacterial Agents; D055767:Immobilized Proteins; D011956:Receptors, Cell Surface; D061905:Single-Domain Antibodies; D054299:env Gene Products, Human Immunodeficiency Virus; C097729:gp140 envelope protein, Human immunodeficiency virus 1; D014640:Vancomycin; D005169:Factor VIII