Characterization for binding complex formation with site-directly immobilized antibodies enhancing detection capability of cardiac troponin I.
The enhanced analytical performances of immunoassays that employed site-directly immobilized antibodies as the capture binders have been functionally characterized in terms of antigen-antibody complex formation on solid surfaces. Three antibody species specific to cardiac troponin I, immunoglobulin G (IgG), Fab, and F(ab')(2) were site-directly biotinylated within the hinge region and then immobilized via a streptavidin-biotin linkage. The new binders were more efficient capture antibodies in the immunoassays compared to randomly bound IgG, particularly, in the low antibody density range. The observed improvements could have resulted from controlled molecular orientation and also from flexibility, offering conditions suitable for binding complex formations.
Cho
Il-Hoon
IH
Program for Bio-Microsystem Technology, Korea University, 1, 5-ka, Anam-dong, Sungbuk-ku, Seoul 136-701, South Korea.
Seo
Sung-Min
SM
Jeon
Jin-Woo
JW
Paek
Se-Hwan
SH
D055910:Antibodies, Immobilized / Q000737:chemistry* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000915:Antibody Affinity; D001710:Biotin / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D007700:Kinetics; D016014:Linear Models; D011485:Protein Binding; D019210:Troponin I / Q000032:analysis* / Q000276:immunology
D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D019210:Troponin I; D001710:Biotin