Immobilization of antibodies on glass surfaces through sugar residues.
This work characterizes substrates for immunoassays obtained through the immobilization of vectorially oriented antibodies on glass. The method of preparation is based on the condensation reaction between an aldehyde group on the F(c) portion of antibodies and the hydrazide group on the modified glass surface. Light microscopy and AFM imaging in height and phase modes were used to assess the properties of the modified surface. Both techniques are consistent with a fairly uniform antibody coverage on the micrometer and submicrometer scales. ELISA tests were used to evaluate the activity and surface distribution of immobilized antibodies as well as nonspecific binding to surfaces after various modification steps. It was shown that exposure of the surfaces to a BSA solution minimized nonspecific binding to undetectable levels.
Gering
John P
JP
Department of Chemistry, Clemson University, Clemson, South Carolina 20634, USA.
Quaroni
Luca
L
Chumanov
George
G
D000906:Antibodies / Q000737:chemistry* / Q000648:ultrastructure; D000937:Antigen-Antibody Reactions; D001665:Binding Sites; D002241:Carbohydrates / Q000737:chemistry*; D005898:Glass / Q000737:chemistry*; D006834:Hydrazines / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology; D018625:Microscopy, Atomic Force; D010084:Oxidation-Reduction; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology; D013499:Surface Properties
D000906:Antibodies; D002241:Carbohydrates; D006834:Hydrazines; D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine