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1.
Daniela Stllner Walter Stcklein Frieder Scheller Axel Warsinke 《Analytica chimica acta》2002,470(2):3438-119
An amperometric immunosensor for hemoglobin-A1c (HbA1c) determination has been developed utilizing membrane-immobilized haptoglobin as affinity matrix fixed in front of a Pt-working electrode. The HbA1c assay was carried out in a two-step procedure including the selective hemoglobin enrichment on the sensor surface and the specific HbA1c detection by a glucose oxidase (GOx) labeled anti-HbA1c antibody. Hydrogen peroxide generated by the enzyme label was oxidized at +600 mV versus Ag/AgCl. A standard curve for HbA1c was obtained with a linear range between 0 and 25% HbA1c of total hemoglobin which correspond to 7.8–39 nM. ELISA studies confirmed the advantage of a sandwich-type format with haptoglobin as capture molecule for selective hemoglobin binding over the direct adsorption method. Results by the sandwich immunoassay showed a linear correlation within the clinically relevant range 5–20% (CV < 3). For sensor application the immobilization procedure of haptoglobin onto CDI-activated cellulose membranes was optimized. 相似文献
2.
Yuan Ruo Cao ShuRui Chai YaQin Gao FengXian Zhao Qing Tang MingYu Tong ZhongQiang Xie Yi 《中国科学B辑(英文版)》2007,50(5):620-628
Alternate adsorption of positively charged colloid-Au nanoparticles (nano-Au⊕) and negatively charged hemoglobin (Hb) on L-cysteine (L-cys) modified gold electrode resulted in the assembly of {Hb/nano-Au⊕}n layer-by-layer films/L-cys modified gold electrode. The nano-Au⊕ was characterized by transmission electron micrograph (TEM) and microelectrophoresis. The modified electrode interface morphology was characterized by electrochemical impedance spectroscopy (EIS), atomic force mi- croscopy (AFM), cyclic voltammograms (CV) and chronoamperometry. Direct electron transfer between hemoglobin and gold electrodes was studied, and the apparent Michaelis-Menten constant ( km app) of the modified electrode was evaluated to be 0.10 mmol·L?1. Moreover, the higher activity of proteins in the nano-Au⊕ films could be retained compared with the electropolymerization membrane, since the pro- teins in nano-Au⊕ films retained their near-native structure. Direct electron transfer between hemoglo- bin and electrode and electrochemically catalyzed reduction of hydrogen peroxide on a modified elec- trode was studied, and the linear range was from 2.1×10-8 to 1.2 ×10?3 mol·L-1 (r = 0.994) with a detection limit of 1.1×10-8 mol·L-1 H2O2. 相似文献
3.
纳米CoHCF修饰电极的制备及其对血红蛋白的电化学测定研究 总被引:1,自引:0,他引:1
A cobalt hexacyanoferrate (CoHCF) nanoparticle (size ca. 60 nm) chemically modified electrode (CME) was fabricated and the electrochemical behavior of hemoglobin (Hb) at this nanosized CoHCF CME was studied. In comparison with a bare glassy carbon electrode (GCE) and a general CoHCF CME electrodeposited in a traditional manner, the present nanosized CoHCF CME performed efficiently electrocatalytic reduction for Hb with relatively high sensitivity, stability, and longlife, Combined with liquid chromatography (LC), the nanosized CoHCF CME was used as the electrochemical detector of Hb in the established flow injection analysis-electrochemical determination (FIA-ECD) system. The peak current was a linear function of concentrations in the range from 2.5×10^-8 to 5.0×10^-6mol/L for Hb, with detection limit of 1.4×10^-8 mol/L. The FIA-ECD system has been successfully applied to assess the Hb content of clinic blood samples with advantages of sensitiveness, speediness, easy control and small sample-consumption. 相似文献
4.
The influences of Triton X-100 on hemoglobin (Hb) behaviors were studied by the methods of UV-Vis spectrum, fluorescence spectrum, HPLC, conductivity, zeta potential and negative-staining transmission electron microscope in Hb/acyclovir/Triton X-100/H2O system. With the increase of Triton X-100 concentration in the system, the percentage of the free acyclovir increased from 58%--63% to 90%--94%. The static quenching constant and the association number of acyclovir to Hb decreased. The fluorescence spectrum, conductivity, zeta potential, fluorescence polarization and negative-staining morphology of Hb tended to recover to those of the original state of Hb in the same concentration of Hb. The interaction between Triton X-100 and Hb is stronger than that between acyclovir and Hb. Most Triton-X-100 was associated with Hb at low Triton X-100 concentration. But the interaction of Triton X-100 with Hb was apparently dominant in high Triton X-100 concentration. The Hb structure was unfolded and finally denatured. 相似文献
5.
Viscous properties of partially hemolyzed erythrocyte suspensions were studied with a cone-plate viscometer. A Casson plot was successfully applied for analysis of the viscous properties. When the degree of hemolysis was increased, Casson viscosity (
c) of the partially hemolyzed erythrocyte suspensions increased, but the substitution of hemoglobin solution with the same O2-carrying capacity of hemolyzed fraction (containing ghost) reduced the
c remarkably. The yield stress (f
c) of the intact erythrocyte suspension increased with increasing hematocrit, but thef
c of partially hemolyzed erythrocyte suspension was dependent on both hematocrit and the degree of hemolysis. The apparent viscosity (
a) of erythrocyte suspension was drastically increased with the slight hemolysis, and became maximum at a certain degree of hemolysis which was dependent on both total hemoglobin concentration and shear rate. Such influence of hemolysis on the
a was also evident in the mixture of erythrocyte suspension and hemoglobin solution. These results suggest that the hemoglobin molecule plays a certain role in erythrocyte-erythrocyte interaction in their suspension, which modifies the viscosity. 相似文献
6.
We have encapsulated several hemoproteins in the sol-gel glass to catalyze the oxidation reaction of dibenzothiophene (model
for organic sulfur compounds in coal) with hydrogen peroxide. In addition to cytochrome c and myoglobin, which have previously
been encapsulated in sol-gel glasses, two other hemoproteins, horseradish peroxidase and bovine blood hemoglobin, have now
been successfully immobilized in sol-gel media with the retention of their spectroscopic properties. All four hemoproteins
studied also demonstrate similar catalytic activities toward the oxidation of dibenzothiophene as compared with the results
obtained with the proteins in solution. In the case of encapsulated cytochrome c, the more water-soluble S-oxide was obtained
with much higher selectivity over the less water-soluble sulfone (S-oxide/sulfone = 7.1) as compared to what was obtained
in the aqueous/organic medium (S-oxide/sulfone = 2.3). Because of the advantage of easy separation of the encapsulated proteins
from the liquid reaction mixture, it is clear from these studies that the immobilization of active hemoproteins in the solid
glass media could serve as more practical biocatalysts. 相似文献
7.
8.
The nitrosyl-hemoglobin (HbNO) is the carrier of nitric oxide (NO) which is the important messenger molecule displaying multiple physiologic and pathophysiologic roles. However it is still not clear for the fate of HbNO molecules during the venous-arterial transit. In this letter, the HbNO transition in vitro was studied by using the electron paramagnetic resonance (EPR) spectra. It was found that HbNO molecules were stable when oxygen did not exist in the system but not stable in aerobic conditions. The absorption spectra further revealed that the methemoglobin (metHb) was the product of HbNO in aerobic environment, showing that the HbNO changed to metHb when there were enough oxygen molecules in the system. 相似文献
9.
0IntroductionStudies of direct electrochemistry of proteins orenzymes at electrodes can serve as a basis for build-ing electrochemical biosensors,enzymatic bioreactors,and biomedical devices[1].This approach simplifiessuch devices without using mediators and is of partic-ular significance for fabricating the third generationbiosensors[2].For example,if a protein or enzyme im-mobilized on electrode surface is capable of directelectron transfer without loss of bioactivities,it can beused in the … 相似文献
10.
Artificial red cells with crosslinked hemoglobin membranes 总被引:1,自引:0,他引:1
Thomas A. Davis William J. Asher Herbert W. Wallace 《Applied biochemistry and biotechnology》1984,10(1-3):123-132
Artificial cells containing concentrated hemoglobin (Hb) solution were prepared by interfacial polymerization of Hb with glutaraldehyde
(GA) in liquid membrane capsules (LMC). A solution containing 30% of Hb was emulsified in mineral oil as red cell-size microdroplets,
and this emulsion was dispersed in an aqueous phase containing glutaraldehyde to form LMC. The LMC were semipermeable templates
that held the microdroplets of Hb in suspension while GA diffused through the oil to the microdroplet surfaces. The GA crosslinked
Hb at the surface of each microdroplet to form an artificial red-cell membrane encapsulating Hb solution. A water-soluble
surfactant was used to eject the cells from the LMC and suspend them in saline.
Several surfactants were evaluated. Cell size was controlled by agitation speed during preparation of the original emulsion.
Cells of 2.52 = ±1.69 μm were prepared. The encapsulated Hb retained capacity to bind and release O2. The cells had aP
50 of 8.9 torr (1200 Pa) and a capacity of 0.55 cc O2/g of total Hb, indicating that the crosslinked portion of the Hb did not contribute to O2 transport. 相似文献