title: Characterization of three amino-functionalized surfaces and evaluation of antibody immobilization for the multiplex detection of tumor markers involved in colorectal cancer.
abstract: Antibody microarrays are powerful and high-throughput tools for screening and identifying tumor markers from small sample volumes of only a few microliters. Optimization of surface chemistry and spotting conditions are crucial parameters to enhance antibodies' immobilization efficiency and to maintain their biological activity. Here, we report the implementation of an antibody microarray for the detection of tumor markers involved in colorectal cancer. Three-dimensional microstructured glass slides were functionalized with three different aminated molecules ((3-aminopropyl)dimethylethoxysilane (APDMES), Jeffamine, and chitosan) varying in their chain length, their amine density, and their hydrophilic/hydrophobic balance. The physicochemical properties of the resulting surfaces were characterized. Antibody immobilization efficiency through physical interaction was studied as a function of surface properties as well as a function of the immobilization conditions. The results show that surface energy, steric hindrance, and pH of spotting buffer have great effects on protein immobilization. Under optimal conditions, biological activities of four immobilized antitumor marker antibodies were evaluated in multiplex immunoassay for the detection of the corresponding tumor markers. Results indicated that the chitosan functionalized surface displayed the highest binding capacity and allowed to retain maximal biological activity of the four tested antibody/antigen systems. Thus, we successfully demonstrated the application of amino-based surface modification for antibody microarrays to efficiently detect tumor markers.
authors.affiliation: Université de Lyon, Institut des Nanotechnologies de Lyon (INL)-UMR CNRS 5270, Ecole Centrale de Lyon, 36 Avenue Guy de Collongue, 69134 Ecully cedex, France.
authors.forename: Zhugen
authors.initials: Z
authors.lastname: Yang
authors.forename: Yann
authors.initials: Y
authors.lastname: Chevolot
authors.forename: Thomas
authors.initials: T
authors.lastname: Géhin
authors.forename: Vincent
authors.initials: V
authors.lastname: Dugas
authors.forename: Nicolas
authors.initials: N
authors.lastname: Xanthopoulos
authors.forename: Vincent
authors.initials: V
authors.lastname: Laporte
authors.forename: Thierry
authors.initials: T
authors.lastname: Delair
authors.forename: Yasemin
authors.initials: Y
authors.lastname: Ataman-Önal
authors.forename: Geneviève
authors.initials: G
authors.lastname: Choquet-Kastylevsky
authors.forename: Eliane
authors.initials: E
authors.lastname: Souteyrand
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Laurenceau
mesh_terms: D000588:Amines / Q000737:chemistry*; D000906:Antibodies / Q000737:chemistry*; D014408:Biomarkers, Tumor / Q000032:analysis*; D048271:Chitosan / Q000737:chemistry*; D015179:Colorectal Neoplasms / Q000175:diagnosis*; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D015394:Molecular Structure; D011437:Propylamines / Q000737:chemistry*; D040081:Protein Array Analysis; D012821:Silanes / Q000737:chemistry*; D013499:Surface Properties
chemical_list: C582385:(3-aminopropyl)dimethylethoxysilane; D000588:Amines; D000906:Antibodies; D014408:Biomarkers, Tumor; D011437:Propylamines; D012821:Silanes; D048271:Chitosan