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81.
《Electroanalysis》2017,29(12):2818-2831
Immobilization of biomolecules with a proper orientation is considered as a basis for diverse biotechnological applications. Herein, we report a host‐guest inclusion complexation between β‐cyclodextrin (β‐CD) and biotin as a versatile approach for the immobilization of biomolecules. As a practical application, a sandwich‐type electrochemical immunosensor was designed for the determination of prostate specific antigen (PSA). The immunosensor was fabricated by in situ electropolymerization of poly(N‐acetylaniline) onto a rGO‐modified Pt electrode. Then, β‐CD was covalently grafted onto the over‐oxidized polymer backbone. For improving the efficiency of the assay, AuNPs were casted on the polymeric film, on the surface of which thionine (TH) as an electron mediator was covalently immobilized. Using a host‐guest inclusion complexation between β‐CD and biotin, a β‐CD/biotin‐Ab1/PSA/Ab2‐horseradish peroxidase (HRP) sandwich was formed on the electrode surface. The analytical signal was produced via electrochemical reduction of THox, generated by biocatalytic oxidation of the THred in the presence of HRP/H2O2. Under optimal conditions, the proposed sensor responded linearly to PSA in the range from 10.0 pg mL−1 to 25.0 ng mL−1, with a low detection limit of 6.7 pg mL−1 (S/N=3). Kinetic parameters of the interaction of β‐CD with Ab1 were also investigated. Finally, the applicability of the immunosensor was successfully investigated for the detection of PSA in human serum samples. 相似文献
82.
Xuecheng Jiang Lixiao Zhou Jie Cheng Hua Zhang Huixia Wang Zhixiong Chen Feng Shi Chenggang Zhu 《Analytica chimica acta》2014
Tumorigenesis is the cumulative result of multiple gene mutations. The mutant proteins that are expressed by mutant genes in cancer cells are secreted into the blood and are useful biomarkers for the early diagnosis of cancer. However, some difficulties exist; for example, the same gene will express different protein mutants in different patients, and early tumors secrete only small amounts of mutant protein. Thus, the presence of mutant proteins in plasma has not previously been exploited for the early diagnosis of cancer. Proximity ligation assay is a protein-detection method that has been developed in recent years and has been widely used because of its high sensitivity. However, this approach still suffers from some shortcomings that should be addressed. In this paper, we develop a covalent-bonding tube-based proximity ligation assay (TB-PLA). The limit of detection of TB-PLA for 0.001 pM, and the method exhibited a broad dynamic range of up to seven orders of magnitude. Furthermore, we coupled the conformation-specific antibody PAb240 of p53 mutants to PCR tubes for TB-PLA. The assay was capable of detecting an approximately 500-fold lower concentration of mutant p53 in serum compared with sandwich ELISA. Thus, we demonstrate TB-PLA to be a highly sensitive and effective approach that is suitable for the early clinical diagnosis of cancer using the conformation-specific antibodies of protein mutants. 相似文献
83.
Satish K. Pandey Aaydha C. Vinayaka Dharam B. Rishi Praveen Rishi C. Raman Suri 《Analytica chimica acta》2014
Typhoid fever is a life threatening bacterial infection that remains a major global health concern. This continued high burden associated with significant morbidity and mortality rate demands specific and rapid detection technique. This work reports a new sandwich type fluorescence immunoassay format using polymyxin B, a cationic receptor molecule, as a binder agent while anti-Vi antibody served as the capturing agent for specifically detecting Salmonella enterica serovar Typhi. Anti-Vi IgG antibody raised against Vi–BSA conjugate revealed affinity of 7.779 nM−1 signifying immunodominancy of O-acetyls groups in Vi polysaccharide. The detection limit of the developed assay was around 101 cells mL−1 of Vi expressing Salmonella enterica serovar Typhi with a correlation coefficient (R2) equal to 0.97. Positive response obtained for all the tested serovar Typhi clinical isolates as well as the pathogen spiked blood samples recommended specificity and accuracy of Vi antigen as a biomarker during typhoid fever. The intra- and inter-assay precision with Vi spiked samples were satisfactory revealing coefficient of variance (CV%) with a mean of 4.05% and 5.97% respectively. This may be the novel attempt and constructive report on the fluorescence based detection of Vi antigen of serovar Typhi in the epidemic as well as pandemic outbreaks. 相似文献
84.
Eichler and Zagier developed a theory of Jacobi forms to understand and extend Maass' work on the Saito‐Kurokawa conjecture. Later Skoruppa introduced skew‐holomorphic Jacobi forms, which play an important role in understanding liftings of modular forms and Jacobi forms. In this paper, we explain a relation between Jacobi forms and skew‐holomorphic Jacobi forms in terms of a group cohomology. More precisely, we introduce an isomorphism from the direct sum of the space of Jacobi cusp forms on and the space of skew‐holomorphic Jacobi cusp forms on with the same half‐integral weight to the Eichler cohomology group of with a coefficient module coming from polynomials. 相似文献
85.
Jackeray R Abid CK Singh G Jain S Chattopadhyaya S Sapra S Shrivastav TG Singh H 《Talanta》2011,84(3):952-962
Present work demonstrates the utilization of surface modified polycarbonate (PC) membrane as solid phase and antibody conjugated CdSe/ZnS quantum dots (QDs) as fluorescent label for the sensitive and selective detection of Salmonella typhi (S. typhi) in water in a period of 2.5 h. PC membrane was surface modified with glycine and activated by EDC/NHS for immobilization of S. typhi specific IgG. Antibody immobilized porous PC membrane was incubated with bacteria contaminated water for immunocapturing of S. typhi. Antibody conjugated QDs were also prepared by using carbodiimide chemistry. Both modified PC membrane and quantum dots were characterized by using various modern analytical tools. It was estimated that 1.95 molecules of QDs were successfully bio-conjugated per unit of IgG. PC membrane with captured bacteria was incubated with prepared IgG conjugated QDs for the formation of sandwich complex. Analysis of the regions of interest (ROI) in fluorescent micrographs showed that newly developed method based on PC and fluorescent QDs has 100 times higher detection sensitivity (100 cells/mL) as compared with detection using conventional dye (FITC) based methods. 相似文献
86.
A new label-free amperometric immunosensor was developed for detection of carcinoembryonic antigen (CEA) based on chitosan-ferrocene (CS-Fc) and nano-TiO2 (CS-Fc + TiO2) complex film and gold nanoparticles-graphene (Au-Gra) nanohybrid. CS-Fc + TiO2 composite membrane was first modified on a bare glass carbon electrode. Then Au-Gra nanohybrid was formed on the CS-Fc + TiO2 membrane by self-assembly strategy. Next, further immobilization of anti-CEA was constructed according to the strong interaction between Au-Gra and the amido groups of anti-CEA. Since Au-Gra nanohybrid films provided a congenial microenvironment for the immobilization of biomolecules, the surface coverage of antibody protein could be enhanced and the sensitivity of the immunosensor has been improved. The good electronic conductive characteristic might be attributed to the synergistic effect of graphene nanosheets and Au NPs. The modified process was characterized by scanning electron microscope (SEM) and cyclic voltammetry (CV). Under optimized conditions, the resulting biosensor displayed good amperometric response to CEA with linear range from 0.01 to 80 ng/mL and a detection limit of 3.4 pg/mL (signal/noise = 3). The results demonstrated that the immunosensor has advantages of high conduction, sensitivity, and long life time. This assay approach showed a great potential in clinical applications and detection of low level proteins. 相似文献
87.
A novel piezoelectric (PZ) immunosensor for the direct detection of malarial Plasmodium falciparum histidine rich protein-2 (PfHRP-2) antigen was developed. The mixed self-assembled monolayers (SAMs) of thioctic acid and 1-dodecanethiol were formed on gold surface of quartz crystal. Cyclic voltammetry, electrochemical impedance spectroscopy and surface Raman spectroscopy techniques were used to characterize the mixed SAMs. The rabbit anti-PfHRP-2 antibodies were coupled on mixed SAM modified gold surface of quartz crystal via NHS/EDC activation method. The PZ immunosensor was applied to detect PfHRP-2 in the linear range of 15-60 ng/ml with a detection limit of 12 ng/ml. It was also found that even after 14 days of storage, 50% of the activity still remained. Clinical human serum samples were tested with this method, and the results were in agreement with those obtained from commercially available ICT kit (NOW® Malaria). 相似文献
88.
89.
SONG ZhongJu YUAN Ruo CHAI YaQin CHE Xin&LV Ping Key Laboratory on Luminescence Real-Time Analysis Ministry of Education College of Chemistry Chemical Engineering Key Laboratory of Eco-environments in Three Gorges Reservoir Region Southwest University Chongqing China 《中国科学B辑(英文版)》2011,(3)
An effective electrochemical signal amplification strategy based on enzyme membrane modification and redox probe immobilization was proposed to construct an amperometric immunosensor.L-cysteine@ferrocene functionalized chitosan,which possessed not only efficient redox-activity but also excellent film-forming ability,was coated on the bare glass carbon electrode. Moreover,the thiol groups(SH)in the ferrocenyl compound were used for gold nanoparticles immobilization via the strong bonding interaction,which co... 相似文献
90.
Li-Ming Yeh 《Journal of Differential Equations》2011,250(4):1828-1849
A priori estimate for non-uniform elliptic equations with periodic boundary conditions is concerned. The domain considered consists of two sub-regions, a connected high permeability region and a disconnected matrix block region with low permeability. Let ? denote the size ratio of one matrix block to the whole domain. It is shown that in the connected high permeability sub-region, the Hölder and the Lipschitz estimates of the non-uniform elliptic solutions are bounded uniformly in ?. But Hölder gradient estimate and Lp estimate of the second order derivatives of the solutions in general are not bounded uniformly in ?. 相似文献