[Immunonanogold catalytic spectrophotometric determination of trace complement 3].
Gold nanoparticles about 10 nm in size were prepared by improved trisodium citrate reduction procedure, and were used to label goat anti-human C3 to obtain a sensitive spectral probe for complement 3 (C3) in the condition of pH 7.5. The immune reaction between nanogold-labeled C3 antibody (anti-C3) and the antigen C3 took place to form the nanogold immune complex in pH 5.6 Na2 HPO4-C6H8O7 buffer solution and in the presence of polyethylene (PEG). The optimal immunoreaction conditions were pH 5.6-9.7 microg x mL(-1) nanogold-labeled anti-C3, 6.0% PEG 6000 and incubation time 15 min under ultrasonic irradiation. After centrifuging for 10 min at 12 000 r x min(-1), the excess nanogold-labeled anti-C3 in the upper solutions was obtained, and was used to catalyze the colored particle reaction between HAuCl4 and NH2 OH x HCl to produce gold particles with bigger size. The influence on the immunonanogold catalytic reaction was considered spectrophotometrically. A pH 2.97 Na3C6H5O7-HCl buffer solution, 53.33 microg x mL(-1) HAuCl4, 74.13 microg x mL(-1) NH2OH x HCl, and reaction time of 3 min at 37 degrees C water bath were chosen for use. Results demonstrated that with increasing C3, the concentration of gold labeled anti-C3 in the upper solution decreased, and the absorbance decreased linearly. Linear relationships between the decreased absorbance and the C3 concentration in the range of 0.025-0.60 ng x mL(-1) were obtained by spectrophotometry at 760 nm. The regress equation was deltaA760 nm = 0.276c+0.025 4, the correlation coefficient was 0.990 3, and the detection limit reached 0.007 2 ng x mL(-1) of C3. The influence of foreign substances such as HAS, BSA, and ammonia acid on the determination of 0.2 ng x mL(-1) of C3was examined. Results showed that this assay has high selectivity. The sensitive, rapid and highly specific assay was applied to the quantification of C3 in human sera, with satisfactory results.
Huang
Wen-Xin
WX
School of Environment and Resource, Guangxi Normal University, Guilin 541004, China.
Jiang
Zhi-Liang
ZL
Liang
Ai-Hui
AH
D002384:Catalysis; D003176:Complement C3 / Q000032:analysis* / Q000276:immunology*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D013053:Spectrophotometry
D003176:Complement C3; D006046:Gold