Increased sensitivity in antigen detection with fluorescent latex nanosphere-IgG antibody conjugates.
IgG antibodies were conjugated to Kodak X-Sight nanospheres to develop fluorescent-labeled antibodies using two different synthetic routes: one involving the DTT reduction method, and the other involving Traut's Reagent modification method. These two methods result in different conjugation efficiencies and different performances in antigen detection. Western blotting shows that the nanosphere-IgG antibody conjugates synthesized using the DTT reduction method are more immunospecific than the conjugates synthesized using Traut's Reagent modification method. In addition, the conjugates synthesized using DTT reduction also show higher antigen detection sensitivity than other commercially available fluorescent-IgG antibody conjugates, including Alexa Fluor, Qdot, and CyDye conjugates.
Ji
Tao
T
Carestream Molecular Imaging, Carestream Health, Inc., New Haven, Connecticut 06511.
Muenker
M Catherine
MC
Papineni
Rao V L
RV
Harder
John W
JW
Vizard
Douglas L
DL
McLaughlin
William E
WE
D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000032:analysis* / Q000276:immunology; D005453:Fluorescence; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology*; D007840:Latex / Q000737:chemistry*; D015394:Molecular Structure; D054118:Nanospheres / Q000737:chemistry*
D000906:Antibodies; D000941:Antigens; D007074:Immunoglobulin G; D007840:Latex