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1.
István Szilágyi László. Horváth Imre Labádi Klara Hernadi István Pálinkó Tamás Kiss 《Central European Journal of Chemistry》2006,4(1):118-134
An imidazolate-bridged copper(II)-zinc(II) complex (Cu(II)-diethylenetriamino-μ-imidazolato-Zn(II)-tris(2-aminoethyl)amine perchlorate (denoted as “Cu,Zn complex”) and a simple copper(II) complex (Cu(II)-tris(2-aminoethyl)
amine chloride (“Cu-tren”) were prepared and immobilised on silica gel (by hydrogen or covalent bonds) and montmorillonite
(by ion exchange). The immobilised substances were characterised by FT-IR spectroscopy and their thermal characteristics were
also studied. The obtained materials were tested in two probe reactions: catalytic oxidation of 3,5-di-tert-butyl catechol (DTBC) (catecholase activity) and the decomposition of hydrogen peroxide (catalase activity). It was found
that the catecholase activity of the Cu,Zn complex increased considerably upon immobilization on silica gel via hydrogen bonds and intercalation by ion exchange among the layers of montmorillonite. The imidazolate-bridged copper(II)-zinc(II)
complex and its immobilised versions were inactive in hydrogen peroxide decomposition. The Cu(II)-tris(2-aminoethyl)amine
chloride complex displayed good catalase activity; however, immobilisation could not improve it. 相似文献
2.
在镀银石英晶片上用压电免疫方法测定了0.5~50mg/L范围内的牛血清白蛋白的浓度,得到线性响应;并较全面地研究了压电晶片的回复性使用问题,以0.2mol/L甘氨酸-盐酸(pH=2.0)溶液洗脱时效果较好 相似文献
3.
ZHANG Jing YANG Ming XU Ze-Li BAO Yong-li WU Yin WANG Yue-zeng MENG Xiang-ying JU Xing-da GU Yang LI Yu-xin 《高等学校化学研究》2006,22(3):308-311
Introduction TheTSP50genehasbeenidentifiedasoneofthe testis specificoncogenes,whichisexpressedathigh levelsinapproximately92%ofhumanbreastcancer samples,makingitanattractivemolecularmarkerand apotentialtargetfordiagnosisandtherapy[1].Itisho mologoustomany… 相似文献
4.
Suzuki Masayasu Watamabe Nobuyuki Tamiya Eiichi Kataoka Tetsuro Tokunaga Tohru Karube Isao 《Applied biochemistry and biotechnology》1987,15(3):191-200
A novel tumor-detection system consisting of complementmediated cytolytic reaction and an image processing system was developed
for the simple and rapid determination of tumor cells.
The present system consists of a CCD image sensor, image memory board, personal computer, and microscope.
When monoclonal antibody 3C4, which is specific to the guinea pig hepatoma L-10, was added to cell suspension, only L-10 cytolysis
occurred. Cytolysis caused a decrease in brightness of the cells observed by phase-contrast microscopy. The phase contrast
image of the cells before cytolysis was converted to a digitalized signal and stored in computer memory. After cytolysis,
a brightness threshold above that of lysed cells was subtracted from the digitalized signal and compared to the signal stored
before reaction.
L-10 cells in mixed cell suspension were determined specifically by the system. Measurement time was only 2 sec and overall
time, including reaction time, was approximately 30 min. Since this method does not require a cell washing process, automation
of the whole system is possible. 相似文献
5.
Sol-gel entrapment of monoclonal anti-atrazine antibodies 总被引:1,自引:0,他引:1
A. Turniansky D. Avnir A. Bronshtein N. Aharonson M. Altstein 《Journal of Sol-Gel Science and Technology》1996,7(1-2):135-143
We report the successful doping of a sol-gel matrix with an antibody, retaining its ability to bind free antigen from an aqueous solution. The particular system described is monoclonal anti-atrazine mouse antibody which was entrapped in SiO2 sol-gel matrices, prepared from tetramethoxysilane by several methods. Atrazine was selected as a model compound for this study, within the framework of the development of immunochemical-based methods for monitoring pesticide residues and other organo-synthetic environmental contaminants. Nanogram quantities of atrazine were applied on SiO2 sol-gel columns doped with this antibody, and the amount of eluted antigen was determined by Enzyme Linked ImmunoSorbent Assay (ELISA). Under appropriate sol-gel-forming conditions, high amounts of atrazine were bound to the sol-gels, ranging between 60% and 91% of the amount applied to the column. The combination of the properties of the sol-gel matrix (e.g., stability, inertness, high porosity, high surface area and optical clarity), together with the selectivity and sensitivity of the antibodies, enable extension of this feasibility study to development of a novel group of immunosensors which could be used for purification, concentration and monitoring of a variety of residues from different sources.Contribution from the Agricultural Research Organization (ARO), Bet Dagan, Israel. No., 1697-E, 1995 series. 相似文献
6.
Eldefrawi ME Azer NL Nath N Anis NA Bangalore MS O'Connell KP Schwartz RP Wright J 《Applied biochemistry and biotechnology》2000,87(1):25-35
An automated flow fluorometer designed for kinetic binding analysis was adapted to develop a solid-phase competitive fluoroimmunoassay
for urinalysis of opiates. The solid phase consisted of polymer beads coated with commercial monoclonal antibodies (MAbs)
raised against morphine. Fluorescein-conjugated morphine (FL-MOR) was used as the fluorescein-labeled hapten. The dissociation
equilibrium constant (K
D
) for the binding of FL-MOR to the anti-MOR MAb was 0.23 nM. The binding of FL-MOR to the anti-MOR MAb reached steady state within minutes and was displaced effectively by morphine
and other opiates. Morphine-3-glucuronide (M3G), the major urinary metabolite of heroin and morphine, competed effectively
with FL-MOR in a concentration-dependent manner for binding to the antimorphine MAb and was therefore used to construct the
calibration curve. The sensitivity of the assay was 0.2 ng/mL for M3G. The assay was effective at concentrations of M3G from
0.2 to 50 ng/mL, with an IC50 of 2 ng/mL. Other opiates and heroin metabolites that showed >50% crossreactivity when present at 1 μg/mL included codeine,
morphine-6-glucuronide, and oxycodone. Methadone showed very low crossreactivity (<5%), which is a benefit for testing in
patients being treated for opiate addictions. The high sensitivity of the assay and the relatively high cutoff value for positive
opiate tests allows very small sample volumes (e.g., in saliva or sweat) to be analyzed. A double-blind comparison using 205
clinical urine samples showed good agreement between this single-step competitive assay and a commercially performed enzyme
multiplied immunoassay technique for the detection of opiates and benzoylecgonine (a metabolite of cocaine). 相似文献
7.
Ellen R. Goldman Igor L. Medintz George P. Anderson Brent L. Iverson George Georgiou Hedi Mattoussi 《Analytica chimica acta》2005,534(1):63-67
We report a simple and versatile approach for the conjugation of luminescent CdSe-ZnS core-shell quantum dots (QDs) to proteins through coordination of engineered C-terminal oligohistidine sequences. Several histidine tail containing proteins were self-assembled onto the QD surface using this method. A recombinant antibody specific for the high explosive 2,4,6-trinitrotoluene (TNT) was conjugated to QDs through a carboxy terminal histidine tail and the bioconjugate used to detect TNT by competitive immunoassay. TNT was detected over the range of 10 μg/ml down to 41 ng/ml using the scFv conjugated to QDs. These results open up the possibility to conjugate luminescent QDs to a whole range of proteins to form QD bioconjugates that can be effectively used in bio-oriented applications, such as sensing, imaging, immunoassay and other diagnostics. 相似文献
8.
Two rapid,sensitive and reliable immunoassay methods,namely competitive indirect enzyme-linked immunosorbent assay(CI- ELISA)and colloidal gold-based immunochromatographic assay(CGIA),were developed to detect ofloxacin(OFL).The linear range of the CI-ELISAwas from 0.5 to 128 ng/mL with a limit of detection(LOD)of 0.35 ng/mL.Good recoveries were obtained in analyzing simulated swine urine samples.The CGIA could accurately estimate OFL at concentrations as low as 10 ng/mL in less than 10 min,and test results were read visually without any instrument. 相似文献
9.
Papaverine (1-(3,4-dimethoxybenzyl)-6,7-dimethoxyisoquinoline, PAP) is a member of the benzylisoquinoline sub-group of the opium alkaloids. It has been widely used for treating diseases like pulmonary arterial embolism and renal or biliary colic. In this paper, a specific conjugate of mono-demethylated papaverine-O-carboxylmethyl ether (MDMPAP-O-CME) and bovine serum albumin (BSA) was synthesized and used as the complete antigen (PAP-BSA), with which we successfully obtained a high-titer anti-PAP polyclonal antibody (pAb) by immunization of rabbits. The anti-PAP pAb showed high affinity to papaverine with an affinity constant (Kaff) of 7.3 × 107 L/mol. With this antibody, we established a sensitive immunochemical method for the determination of papaverine based on indirect competitive enzyme-linked immunosorbent assay (ELISA). The optimal concentrations of the coated antigen (PAP-OVA) and purified pAb used in the ELISA were 5 and 1.2 μg/mL, respectively. The cross reactivity of other benzylisoquinoline derived substances, including 1-(3,4-dihydroxybenzyl)-7-hydroxy-6-methoxy-isoquinoline (6-methoxy-papaveroline, MPAPO), morphine (MP) and codeine (CD) were all lower than 1%. The linear range of the calibration curve was 0.1-1000 ng/mL. Normal human serum samples were spiked with known amount of papaverine and measured by the ELISA. Recoveries were between 102% and 105%. Papaverine content in a commercial papaverine hydrochloride injection sample was also determined using the established ELISA. Compared with the results given by the control experiment of HPLC, the recoveries of ELISA to detect injection samples were 102-110%. The limits of detection for synthetic serum samples and injection samples of papaverine hydrochloride were 0.25 and 0.06 ng/mL, respectively. 相似文献
10.
Changes in binding affinity to catalytic antibody 6D9 of chloramphenicol phosphonate derivatives (CPDs) containing H or F were investigated by performing free energy calculations based on molecular dynamics simulations. We calculated the binding free energy, enthalpy, and entropy changes (DeltaDeltaG, DeltaDeltaH, and -TDeltaDeltaS) attributable to H-->F substitution by comparing results for CPDs containing a trifluoroacetylamino group (CPD-F) or an acetylamino group (CPD-H). The calculated DeltaDeltaG, DeltaDeltaH, and -TDeltaDeltaS values were -2.9, -6.3, and 3.5 kcal mol(-1) and close to experimental values observed for a series of similar ligands, chloramphenicol phosphonates with F and H (-1.4, -3.5, and 2.1 kcal mol(-1)). Therefore, CPD-F binds more strongly to 6D9 than does CPD-H. To clarify the origin of the large difference in DeltaDeltaG, we apportioned the calculated values of DeltaDeltaG and DeltaG for the associated and dissociated states into contributions from various atomic interactions. We found that the H-->F substitution increased the binding affinity mainly by decreasing the hydration free energy and not by increasing favorable interactions with the antibody. The decreased hydration free energy of the ligand was mainly due to unfavorable coulombic interactions between the trifluoroacetylamino group and solvent waters, which increased the free energy of the dissociated state (by about 3.7 kcal mol(-1)). Also, the trifluoroacetylamino group slightly increased the free energy level of the associated state (about 0.8 kcal mol(-1)) because favorable van der Waals interactions compensated for unfavorable coulombic interactions with antibody atoms. In addition, the enthalpy and entropy changes, DeltaDeltaH and -TDeltaDeltaS (computationally -6.3 and 3.5 kcal mol(-1)), originated mainly from a decrease in hydration free energy in the dissociated state. The CPD-F and CPD-H ligands had substantially different structures in the dissociated and complexed states. 相似文献