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81.
以琼脂凝胶固定热聚IgG制备类风湿关节炎免疫吸附剂的研究 总被引:2,自引:0,他引:2
以环氧氯丙烷对琼脂凝胶珠进行活化反应后键联热聚IgG,制成一种新型类风湿关节炎免疫吸附剂。确定了最佳制备条件,使凝胶上环氧基的含量达110μmol/g,对热苯IgG的固定量达6mg/g。在体外条件下吸附剂对三种类风湿因子IgMRF,IgGRF及IgARF的吸附量分别达3400,2250和2400IU/g,具有良好的应用前景。 相似文献
82.
Joe Sarobe Iris Miraballes Jos Antonio Molina Jacqueline Forcada Roque Hidalgo-lvarez 《先进技术聚合物》1996,7(9):749-753
Monodisperse core–shell stable latexes with reactive methylchloride surface functionalities were prepared at two different reaction temperatures. The reaction temperature played an important role in the amount of reactive functional groups. The covalent coupling had an efficiency of more than 50%. Antibodies covalently bound to functionalized polystyrene beads were used to detect corresponding antigens by nephelometry. 相似文献
83.
以寡聚组氨酸为配基的高效亲和色谱研究 总被引:2,自引:0,他引:2
亲和色谱在所有色谱分离中选择性最高.以组氨酸为配基的亲和介质(Affinitymedia)对免疫球蛋白IgG具有亲和力,显示了良好的分离效果[1];以组氨酸为配基的亲和色谱分离纯化IgG的机理[2]以及组氨酸亲和色谱分离IgG亚基也已有报道[3].组氨酸配基亲和色谱法具有简单、有效和价廉等特点,在免疫化学、临床分析及血液制品的生产等方面均有良好的应用前景.本文分别以含1,3,5个组氨酸残基的小肽为配基,合成了系列亲和介质,并对其亲和色谱行为进行了评价,研究了不同链长的寡聚组氨酸与IgG的亲和相互作用.1 实验部分1.1 固相肽合成 采用Fmoc肽合… 相似文献
84.
85.
核壳型Fe_3O_4/Au粒子的制备及其在表面等离子体子共振传感器中的应用 总被引:1,自引:0,他引:1
合成了核壳型Fe3O4/Au复合粒子,并对其形貌、光学性质进行了表征.通过外加磁场将Fe3O4/Au复合粒子与兔抗人IgG的偶联体固定于表面等离子体子共振(SPR)传感器的金基底膜上,形成了Fe3O4/Au/抗IgG敏感膜.与传统的通过巯基丙酸连接蛋白的方式相比,磁场作用固定的Fe3O4/Au/抗IgG敏感膜制备简单,易洗脱,具有良好的再生性,且在一定程度上提高了传感器的灵敏度.并对人IgG进行了测定,结果表明,传感器对于浓度范围在1.25~20.00μg·mL-1的人IgG有良好的信号响应. 相似文献
86.
Accurate determination of concentration of immunoglobulin (IgG) to tetanus toxoid is important in order to evaluate the immunogenicity of tetanus toxoid vaccines, immune competence in individual patients and to measure the prevalence of immunity in populations. Surface modified polyacrylonitrile (PAN) fibers were evaluated as a matrix to develop highly sensitive method for the detection of anti-tetanus antibody in a sandwich ELISA format. In the proposed method tetanus toxoid immobilized on modified PAN fibers was used to detect anti-tetanus antibody (raised in horse hence represented as horse anti-tetanus toxoid or HAT-Ab) with horse raddish peroxidase enzyme conjugated with Rabbit anti-Horse IgG (RAH-HRP) as the label within 2.5 h. A sigmoidal pattern for the detection of different concentration of antibody ranging from 1.0 to 0.0001 IU mL−1 was validated. The immunoassay recorded a very high sensitivity as concentration as low as 0.0005 IU mL−1 of HAT-Ab was detected. The intra- and inter-assay precision for 3 parallel measurements of 0.01 and for 0.001 IU mL−1 of antibody varied from 5.4% to 11% and 5.7% to 20% respectively. PAN fibers were also used to qualitatively access the presence of different level of anti-tetanus antibody spiked in human blood. Seroepidemiological studies to measure the immunity against tetanus were conducted with twenty-five human beings belonging to various age groups using modified PAN-ELISA. The sensitivity, specificity and the reproducibility of the developed immunoassay indicate the potential application of modified PAN fibers in the field of immunodiagnostics. 相似文献
87.
The aim of this work was to devise a one‐step purification procedure for monoclonal antibodies (MAbs) of IgG class by immobilized metal affinity chromatography (IMAC). Therefore, several stationary phases were prepared containing immobilized metal chelates in order to study the chromatographic behaviour of MAbs against wild‐type amidase from Pseudomonas aeruginosa. Such MAbs adsorbed to Cu(II), Ni(II), Zn(II) and Co(II)–IDA agarose columns. The increase in ligand concentration and the use of longer spacer arms and higher pH values resulted in higher adsorption of MAbs into immobilized metal chelates. The dynamic binding capacity and the maximum binding capacity were 1.33 ± 0.015 and 3.214 ± 0.021 mg IgG/mL of sedimented commercial matrix, respectively. A KD of 4.53 × 10−7 m was obtained from batch isotherm measurements. The combination of tailor‐made stationary phases of IMAC and the correct selection of adsorption conditions permitted a one‐step purification procedure to be devised for MAbs of IgG class. Culture supernatants containing MAbs were purified by IMAC on commercial‐Zn(II) and EPI‐30–IDA–Zn(II) Sepharose 6B columns and by affinity chromatography on Protein A‐Sepharose CL‐4B. This MAb preparation revealed on SDS–PAGE two protein bands with Mr of 50 and 22 kDa corresponding to the heavy and light chains, respectively. Copyright © 2011 John Wiley & Sons, Ltd. 相似文献
88.
Moreau T Faye C Baqué M Desvignes I Coussot G Pascal R Vandenabeele-Trambouze O 《Analytica chimica acta》2011,(2):354-360
Finding a general solution for optimizing the grafting of antibody on solid surfaces is difficult due to the variety of material, grafting principles and chemistries or surface formats available (beads, microplates, fibers, etc.). Pre-screening methods able to assess grafting efficiency (GE) and specific activity (SA) are required. In this context, we present here two colorimetric assays that can be used on a wide variety of surface format, chemistry, etc. The first one, ADECA (Amino Density Estimation by Colorimetric Assay) allows a rapid estimation of grafted antibodies and allows calculating the GE. The second one, A2HRP (Antibody Anti-HorseRadish Peroxidase) provides a measure of the amount of active antibody, which, combined to ADECA, is used to determine the SA of grafted antibody. Analytical parameters (limit of detection, repeatability, linearity, etc.) of these two colorimetric assays are presented. Using two commercially available microplates, we demonstrated that, when used in parallel, these rapid and sensitive methods are well adapted to pre-screening of antibody grafting performances. 相似文献
89.
Teles FS 《Analytica chimica acta》2011,(1):28-42
The past decades have witnessed enormous technological improvements towards the development of simple, cost-effective and accurate rapid diagnostic tests for detection and identification of infectious pathogens. Among them is dengue virus, the etiologic agent of the mosquito-borne dengue disease, one of the most important emerging infectious pathologies of nowadays. Dengue fever may cause potentially deadly hemorrhagic symptoms and is endemic in the tropical and sub-tropical world, being also a serious threat to temperate countries in the developed world. Effective diagnostics for dengue should be able to discriminate among the four antigenically related dengue serotypes and fulfill the requirements for successful decentralized (point-of-care) testing in the harsh environmental conditions found in most tropical regions. The accurate identification of circulating serotypes is crucial for the successful implementation of vector control programs based on reliable epidemiological predictions. This paper briefly summarizes the limitations of the main conventional techniques for biomolecular diagnosis of dengue disease and critically reviews some of the most relevant biosensors and rapid diagnostic tests developed, implemented and reported so far for point-of-care testing of dengue infections. The invaluable contributions of microfluidics and nanotechnology encompass the whole paper, while evaluation concerns of rapid diagnostic tests and foreseen technological improvements in this field are also overviewed for the diagnosis of dengue and other infectious and tropical diseases as well. 相似文献
90.
A novel type of poly(N-isopropylacrylamide) grafted E-51 epoxy-based monoliths in a 100 mm × 4.6 mm I.D. stainless steel column with well-controlled three-dimensional skeletal structures has been prepared and proposed for the separation of proteins. The grafted PNIPAAm chain via surface-initiated atom transfer radical polymerization was successfully performed. The proposed method provided a new route to modify the E-51 epoxy-based monoliths for widening their applications. Meanwhile, the temperature and the salt concentration responses of the grafted monolithic columns were investigated. Under the salt gradient, six proteins were well separated in hydrophobic interaction mode. Moreover, for further confirming the application of the prepared monolith was meaningful for proteome analysis in actual system, the separation of human serum sample was performed. 相似文献