Detection of label-free cancer biomarkers using nickel nanoislands and quartz crystal microbalance.
We present a technique for the label-free detection and recognition of cancer biomarkers using metal nanoislands intended to be integrated in a novel type of nanobiosensor. His-tagged (scFv)-F7N1N2 is the antibody fragment which is directly immobilized, by coordinative bonds, onto ~5 nm nickel islands, then deposited on the surface of a quartz crystal of a quartz crystal microbalance (QCM) to validate the technique. Biomarker GTPase RhoA was investigated because it has been found to be overexpressed in various tumors and because we have recently isolated and characterized a new conformational scFv which selectively recognizes the active form of RhoA. We implemented a surface chemistry involving an antibiofouling coating of polyethylene glycol silane (PEG-silane) (<2 nm thick) and Ni nanoislands to reach a label-free detection of the active antigen conformation of RhoA, at various concentrations. The methodology proposed here proves the viability of the concept by using Ni nanoislands as an anchoring surface layer enabling the detection of a specific conformation of a protein, identified as a potential cancer biomarker. Hence, this novel methodology can be transferred to a nanobiosensor to detect, at lower time consumption and with high sensitivity, specific biomolecules.
Martínez-Rivas
Adrián
A
LAAS-CNRS, Université de Toulouse, Toulouse, France. nanobiomex@hotmail.com
Chinestra
Patrick
P
Favre
Gilles
G
Pinaud
Sébastien
S
Séverac
Childérick
C
Faye
Jean-Charles
JC
Vieu
Christophe
C
D055910:Antibodies, Immobilized; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000276:immunology; D015374:Biosensing Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D066298:In Vitro Techniques; D053768:Metal Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D050997:Nanomedicine; D009532:Nickel; D011092:Polyethylene Glycols; D058525:Quartz Crystal Microbalance Techniques; D012821:Silanes; D013499:Surface Properties; D020742:rhoA GTP-Binding Protein / Q000032:analysis / Q000276:immunology
D055910:Antibodies, Immobilized; D014408:Biomarkers, Tumor; D012821:Silanes; C490327:poly(ethylene glycol)silane; C066737:RHOA protein, human; D011092:Polyethylene Glycols; D009532:Nickel; D020742:rhoA GTP-Binding Protein