Biochemical and immunochemical characterization of the antigen-antibody reaction on a non-toxic biomimetic interface immobilized red blood cells of crucian carp and gold nanoparticles.
A special protein assay system based on a highly hydrophilic, non-toxic and conductive biominetic interface has been demonstrated. To fabricate such assay system, red blood cells of crucian carp (RBC) was initially grown on a glassy carbon electrode surface (GCE) deposited nano-sized gold particles (GPs), a second gold nanoparticle layer (NG) was then absorbed on the RBC surface, and finally mammary cancer 15-3 antibody (anti-CA15-3) was attached on the functional RBC surface. A competitive immunoassay format was employed to detect CA15-3 with horseradish peroxidase (HRP)-labeled CA15-3 as tracer and hydrogen peroxide as enzyme substrate. When the immunosensor was incubated into a mixture solution containing HRP-labeled CA15-3 and CA15-3 sample for 1h at 37 degrees C, the amperometric response decreased with the increment of CA15-3 sample concentration. AFM images of the modified layer revealed a uniform distribution of protein and nanogold. In situ QCM and electrochemical measurements demonstrated that the wanted antibody-antigen reactions should occur with high specificity and selectivity. The specific immunoassay system can be developed further to yield sophisticated structures for other proteins.
Tang
Dianping
D
College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
Yuan
Ruo
R
Chai
Yaqin
Y
D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis* / Q000276:immunology; D000937:Antigen-Antibody Reactions / Q000276:immunology; D001681:Biological Assay / Q000379:methods*; D040761:Biomimetic Materials / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D002347:Carps; D002478:Cells, Cultured; D018914:Cells, Immobilized; D004563:Electrochemistry / Q000379:methods; D004912:Erythrocytes / Q000276:immunology*; D006046:Gold / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure
D000936:Antigen-Antibody Complex; D006046:Gold