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1.
The immobilization of proteins, especially receptor proteins commonly used in high through-put screening of drugs (HTS), have received great attention in recent years. There are many successful isothermal models for describing the adsorption of protein onto solid surface, such as Langmuir model, Bi-Langmuir model, Fowler model, Freundlich model, Freundlich-Langmuir model and Tekmin model etc. In all these models, Langmuir model was the most favorable one model accepted by many researchers, but the experimental results showed that it was not entirely fit to all adsorption behaviors. So new models were required for describing protein adsorption onto microspheres in different conditions.In our research, a novel isothermal model, including Langmuir and other adsorbing behaviors was presented basing on the holding degree of surface active sites and the interaction styles of protein immobilization. In Langmuir model, the adsorbing amount of protein was described as [PS] =Km[P]/1 + K[P], where [PS] was the concentration of adsorbed protein, [P] was the concentration of freeprotein at equilibrium state, and Km and K was constant. According to the interactions of protein and ligands, there were three patterns in the interactions of protein and ligands. On the similar assumption that the interaction of protein and microspheres were three styles, and based on the definition of the holding degree of surface active sites (Y), three adsorption behaviors could be described as Y K[ P ]φ/ K[P]φ+1 or ln K + φ ln[P] =ln(Y/1-Y) in which [P] was the concentration of free protein at equilibrium state, and φ and K was constant. Different scale of φ presented different adsorption behaviors, especially when φ was 1, the adsorption behavior was Langmuir adsorbing model. Figure I indicated the different adsorbing results in different adsorption behaviors (φ>1, φ<1,and φ=1). 相似文献
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4.
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
5.
A. W. Flounders D. L. Brandon A. H. Bates 《Applied biochemistry and biotechnology》1995,50(3):265-284
Monoclonal antibodies specific for thiabendazole were immobilized to silicon, silicon dioxide, stoichiometric silicon nitride,
and silicon-rich silicon nitride surfaces. This work provides the foundation for the development of a homogeneous sensor system
for rapid detection and quantification of thiabendazole residues in produce and animal tissue. Immobilization was performed
via aqueous silanization of the substrate followed sequentially by treatment with glutaraldehyde and contact with antibody
solution in the presence of detergent. Surfaces were challenged with thiabendazole-horseradish peroxidase conjugate in an
ELISA format to estimate immobilized antibody load. A stable and reproducible surface loading of 2 x 1011 antibodies/cm2 was obtained only after surfaces received postimmobilization treatments to remove nonspecifically adsorbed antibody. No difference
in surface loading was noted when using 30% hydrogen peroxide rather than nitric acid for silanol activation. Little difference
was noted among the antibody loadings achieved on the various silicon substrates. Bound antigen-enzyme conjugate was eluted
with 0.1N acetic acid and reproducible surface activity was measured for up to four consecutive antigen challenges. Immobilized
antibody surfaces were stabilized with 2% sucrose, dehydrated at 37‡C and stored in vacuum or stored at 4‡C in phosphate buffered
saline containing 0.01% sodium azide without significant loss of activity. 相似文献
6.
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
7.
This study investigates the effect of surface immobilization on the bactericidal function of a quaternary ammonium compound. Quaternary ammonium silane (QAS) coated planar surfaces did not produce any measurable mortality of Staphylococcus aureus, while 1 µm QAS‐coated microparticles did produce S. aureus mortality. The experiments using QAS‐coated microparticles indicate that the ability of QAS molecules to disrupt the cell wall is not hindered by covalent immobilization of QAS to a surface. These results provide evidence that S. aureus cells on a QAS‐coated planar surface are not exposed to a sufficient number of QAS molecules to produce significant mortality. This result has important implications for the development of self‐decontaminating coatings. Covalent immobilization is used to prevent leaching of the bactericidal compound. However, covalent immobilization may result in a significant tradeoff in bactericidal performance. Published in 2007 by John Wiley & Sons, Ltd. 相似文献
8.
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
9.
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
10.
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