磺胺二甲嘧啶人工抗原的合成与鉴定

采用重氮化法将磺胺二甲嘧啶(SM2)与牛血清白蛋白(BSA)和卵清蛋白(OVA)交联合成人工抗原SM2-BSA与SM2-OVA,并通过紫外扫描、SDS-PAGE电泳、凝胶层析和红外光谱等方法扫描进一步检测合成效果.紫外扫描检测结果显示,SM2与BSA、OVA偶联成功,SM2-BSA与SM2-OVA的特征波长分别为2762、77 nm;经计算SM2-BSA与SM2-OVA的分子结合比例分别为5.3∶1和18.4∶1;SDS-PAGE电泳迁移率、凝胶层析出峰时间和红外光谱曲线特征均显示SM2与BSA、OVA偶联成功.     

拟除虫菊酯类农药单克隆抗体的制备及鉴定

以间苯氧基苯甲酸(PBA)为半抗原,与牛血清白蛋白(BSA)偶联后作为抗原,免疫Balb/c小鼠,取小鼠脾细胞与骨髓瘤细胞(SP2/0)融合,使用PBA与鸡卵清白蛋白(OVA)偶联物作为包被物,间接ELISA法筛选到4株特异性针对PBA的杂交瘤。其中两株细胞2F7和7B1制备腹水并用ProteinG柱纯化抗体,抗体效价均达到1:625000。单抗7B1用于间接竞争ELISA法,PBA检测下限为0.025μg·mL-1,IC50为0.57μg·mL-1,对溴氰菊酯、甲体氯氰菊酯、甲氰菊酯、氰戊菊酯均有特异性识别,IC50分别为0.79μg·mL-1、0.74μg·mL-1、0.63μg·mL-1、0.8μg·mL-1,对联苯菊酯识别弱,IC50为99.7μg·mL-1。该单抗可用于多种菊酯类农药的检测。     

水相合成 CdS纳米晶标记牛血清白蛋白

利用水相中直接合成的CdS纳米晶，与牛血清白蛋白(BSA)进行偶连标记。通过分子筛层析对标记后的牛血清白蛋白进行纯化，在紫外灯下即可观察到标记蛋白的荧光。对CdS纳米晶标记后的牛血清白蛋白的荧光光谱的研究表明，标记蛋白后的CdS纳米晶其荧光无明显淬灭。     

苏丹红Ⅰ号多克隆抗体的制备与特异性鉴定

采用琥珀酸酐法,分别以BSA、OVA为载体蛋白合成了免疫抗原和包被抗原并通过SDS-PAGE凝胶电泳和紫外扫描法鉴定了合成的抗原为载体蛋白和苏丹红I号的复合物.将合成的免疫抗原用弗氏佐剂乳化后免疫新西兰大白兔,制备出多克隆抗体,经辛酸纯化,间接ELISA法测得有高效价血清抗体;进一步采用琼脂双扩散试验和免疫金试纸法鉴定出血清中含有苏丹红Ⅰ号特异性抗体.     

壳聚糖与牛血清白蛋白分子印迹聚合物的制备与表征

利用壳聚糖为功能单体,在模板分子牛血清白蛋白存在下,采用滴加成球法制备出对牛血清白蛋白(BSA)具有特异识别性能的分子印迹聚合物,并用原子力显微镜(AFM)及红外光谱进行了表征。实验结果表明,利用壳聚糖制备的牛血清白蛋白分子印迹聚合物对牛血清白蛋白具有特异识别功能,显示出明显的印迹效果。这种特异分子印迹介质的制备,为分离提纯特种结合蛋白提供了一种优良方法。     

两步法制备两性离子陶瓷复合膜及抗污染研究

为了减弱蛋白与陶瓷膜表面的作用力,采用两步表面接枝法,首先采用硅烷化反应将叔胺氢给体(N,N-二甲基-3-氨丙基)三甲氧基硅烷(DMAPS)偶联到含有羟基的陶瓷膜表面,再引发2-溴乙基磺酸钠(SBTS)在膜表面发生季铵化反应,制备磺酸型两性离子陶瓷复合膜(SBTS-c-DMAPS-g-Al_2O_3).结果表明:红外光谱测试显示,与未改性膜表面相比,改性膜表面于1 065cm~(-1)和1 200cm~(-1)处分别出现C—N峰与SO3-峰.接触角测试结果表明,膜的亲水性得到改善,膜表面水滴接触角由36°减小到21°,纯水通量增大20%,可间接表明材料表面成功接枝上两性离子.牛血清白蛋白(BSA)的动态过滤实验表明,改性膜对BSA溶液渗透率比未改性膜的BSA溶液渗透率高30%,改性膜被污染后,再用纯水清洗后的纯水通量恢复率达到90%,而未改性膜被污染后,再用纯水清洗后的纯水通量恢复率只有26%.这是因为两性离子陶瓷复合膜具有易清洗性能.     

磁性荧光纳米复合粒子的制备及其表面生物功能化

用化学共沉淀法制备了四氧化三铁(Fe3O4)纳米粒子,Stober法在其表面包裹SiO2并复合荧光标记物FITC,EDC偶联牛血清白蛋白(BSA)。采用扫描电子显微镜(SEM),傅里叶变换红外光谱仪(FT-IR),荧光分析仪,电子能谱(XPS)等对复合粒子进行了表征。结果表明,复合粒子保留了FITC的荧光特性、Fe3O4的磁响应性并成功接枝蛋白分子。生物功能化磁性荧光复合纳米粒子有望广泛应用于细胞标记、荧光追踪、磁性分离等领域。     

壳聚糖胃滞留-漂浮微球体外释药的高精度组合建模法研究

以牛血清白蛋白(BSA)为模型药物,壳聚糖(CS)和海藻酸钠(SA)为壁材,采用锐孔凝固浴法制备了壳聚糖胃滞留-漂浮微球(CGRM).建立零级释放动力学模型(ZODM)、Higuchi模型和Ritger-Peppas(RPcppas)模型对CGRM的释药性能进行了模拟和预测研究,并在此基础上构建了一种高精度的组合建模法.组合模型的模拟和预测精度均有一定提高,表明该组合建模法是一种模拟长效药物缓释体系释药规律的有力工具.     

近期期刊文摘

新型磷-氮阻燃剂的合成及其与三聚氰胺氰脲酸盐在PA6中的协效应用工程塑料应用2006(11),9合成了一种新型磷-氮阻燃剂N,N‘-哌嗪四苯氧基氨基磷酸酯(PTPP),将PTPP与三聚氰胺氰脲酸盐(MC)的复合物添加到聚     

碳、铈共掺杂改性TiO2负载ACFS复合水净化材料的制备与性能

分别采用偶联法与溶胶-凝胶法制备了碳、铈共掺杂改性TiO<,2>负载活性碳纤维(ACFS)复合水净化材料(Ce,C-TiO<,2>/ACFS).以亚甲基蓝模拟废水的脱色效果为考察目标,对其负载量进行了优化,得出最佳负载率分别为:5.82%和8.41%.扫描电镜(SEM)表征显示:偶联法制备的复合材料中,改性TiO<,2...     

Efficient immobilization and patterning of biomolecules on poly(ethylene terephthalate) films functionalized by ion irradiation for biosensor applications

The surface of a poly(ethylene terephthalate) (PET) film was selectively irradiated with proton beams at various fluences to generate carboxylic acid groups on the surface; the resulting functionalized PET surface was then characterized in terms of its wettability, chemical structure, and chemical composition. The results revealed that (i) carboxylic acid groups were successfully generated in the irradiated regions of the PET surface, and (ii) their relative amounts were dependent on the fluence. A capture biomolecule, anthrax toxin probe DNA, was selectively immobilized on the irradiated regions on the PET surface. Cy3-labeled DNA as a target biomolecule was then hybridized with the probe DNA immobilized on the PET surface. Liver-cancer-specific α-fetoprotein (AFP) antigen, as a target biomolecule, was also selectively immobilized on the irradiated regions on the PET surface. Texas Red-labeled secondary antibody was then reacted with an AFP-specific primary antibody prebound to the AFP antigen on the PET surface for the detection of the target antigen, using an indirect immunoassay method. The results revealed that (i) well-defined micropatterns of biomolecules were successfully formed on the functionalized PET surfaces and (ii) the fluorescence intensity of the micropatterns was dependent mainly on the concentrations of the target DNA hybridized to the probe DNA and the target AFP antigen immobilized on the PET films. The lowest detectable concentrations of the target DNA and target AFP antigen in this study were determined to be 4 and 16 ng/mL, respectively, with the PET film prepared at a fluence of 5 × 10(14) ions/cm(2).     

Immobilization of proteins in immunochemical microarrays fabricated by electrospray deposition.

Electrospray (ES) deposition has been applied to fabricate protein microarrays for immunochemical assay. Protein antigens were deposited as arrays of dry spots on a surface of aluminized plastic. Deposition was performed from water solutions containing a 10-fold (w/w of dry protein) excess of sucrose. Upon contact with humid air, the spots turn into microdroplets of sucrose/protein solution from which proteins were either adsorbed or covalently linked to clean or modified aluminum surfaces. It was found that covalent binding of antigens via aldehyde groups of oxidized branched dextran followed by reduction of the Schiff bonds gives the highest sensitivity and the lowest background in microarray-based ELISA, as compared to other tested methods of antigen immobilization. The minimum concentration of a primary mouse antibody detected in indirect ELISA with such antigen microarrays was approximately 0.3-1.0 ng/mL for ELF-97 or BCIP/NBT substrates of alkaline phosphatase.     

SERS biodetection using gold-silica nanoshells and nitrocellulose membranes

We have developed a rapid, reproducible, easy to execute, surface enhanced Raman scattering (SERS) method for detection of low volumes and total amounts of biological antigens using an analyte capture system derived from methods commonly used in Western blotting. Our method is a "half-sandwich" assay with an antigen detection scheme that employs a nitrocellulose (NC) membrane with 200 nm pore size to capture subnanograms of analyte and concentrate them in a small area from applied volumes as low as one microliter. The SERS probes used for detection consist of gold-silica nanoshells modified with a two-component mixed monolayer system. One component consists of a poly(ethylene glycol) (PEG)-modified Raman-active chromophore bound to the gold surface which allows for SERS detection and imparts particle stability. The second component uses (ortho-pyridyl) disulfide-PEG-succinimidyl ester to couple the recognition antibody to the particle surface. By controlling the reaction time and concentration of thiols, a mixed monolayer is prepared on the nanoshell surface with the ability to recognize low concentrations of analyte and generate reproducible SERS signals. Using this strategy, we have achieved SERS signals that are proportional to antigen present on the membrane allowing detection of total antigen amounts as low as 1.25 ng for some cases. The performance of this new SERS bioassay has been tested with a variety of potential antigens, demonstrating the potential for multiplexed detection of analytes.     

A Knowledge-Based Pilot Study on Assessing the Music Influence

A knowledge-driven approach is proposed for assessing the music influence on university students. The proposed method of modeling and conducting the interactive pilot study can be useful to convey other surveys, interviews, and experiments created in various phases of the user interface (UI) design processes, as part of a general human-computer interaction (HCI) methodology. Benefiting from existing semantic Web and linked data standards, best practices, and tools, a microservice-oriented system is developed as a testbed platform able to generate playlists in a smart way according to users’ music preferences. This novel approach could bring also benefits for user interface adaptation by using semantic Web techniques. Statistical analysis based on the ANOVA method and post-experiment survey data led to the conclusion that music listened has a significant impact on students’ cognitive abilities in various contexts. All obtained results were semantically enhanced by using different conceptual models in order to create a knowledge graph providing support for automated reasoning. Also, a knowledge-based persona Web smart editor was implemented in order to include music preferences for certain categories of the potential users operating a specific interactive system.     

Time-resolved fluorescence detection of mosaic DNA chip

We demonstrated a time-resolved fluorescence (TRF) label and detection of mosaic DNA chip in this paper. We synthesized oligonucleotide sequences in situ on glass slides directly, and then sliced them up into small pieces and patched up the pieces bearing different sequences to generate a mosaic DNA chip. With multiple 4, 7-bis(chlorosulfophenyl)-1, 10-phenanthroline-2, 9-dicarboxylic acid (BCPDA, abbreviated as BCPDA) labeling method based on avidin-biotin amplification, we established a TRF detection format on the mosaic DNA chip. The detection method allows discriminatory signals for perfect match, one-base mismatch, two-base mismatch, and three-base mismatch by TRF labeled DNA hybridization, whereby Europium (III, Eu3+) was captured and released on the principle of complexation and dissociation interaction between BCPDA and Eu3+ solution when the BCPDA-tagged avidin and biotin-capped oligonucleotide sequence linked. The fluorescence spectra and related lifetimes were determined. We also compared the TRF detection mode with the conventional fluorescence one. These results showed the former is a potential alternative replacement of the latter, especially for labeling the mosaic DNA chip. The discovery is of fundamental interest and has significant implications to biochips and biosensors based on time-resolved-fluorescence detection.     

Latex of immunodiagnosis for detecting the Chagas disease: II. Chemical coupling of antigen Ag36 onto carboxylated latexes

A novel immunodiagnosis reagent for detecting the Chagas Disease was developed, by chemical coupling of antigen Ag36 of Trypanosoma cruzi onto two (carboxylated and core-shell) latexes. The coupling reactions involved the use of a carbodiimide intermediate. Bovine serum albumin (BSA) was used as a model protein for determining the appropriate conditions for its physical and chemical coupling. BSA showed an increased adsorption onto the base carboxylated latexes, with respect to a PS latex without carboxyl groups. The chemical bonding experiments only involved the carboxylated latexes. With BSA, the final density of covalently bound protein was 2.30 mg/m². In addition, around 55% of the total linked protein was chemically coupled, and the reaction was little affected by the pH. With Ag36, the final density of covalently bound protein was 2.44 mg/m², around 80% of the total linked protein was chemically coupled, and the chemical coupling was maximum at pH = 5 (i.e., close to the isoelectric point).     

Alpha-alumina nanoparticles induce efficient autophagy-dependent cross-presentation and potent antitumour response

Therapeutic cancer vaccination is an attractive strategy because it induces T cells of the immune system to recognize and kill tumour cells in cancer patients. However, it remains difficult to generate large numbers of T cells that can recognize the antigens on cancer cells using conventional vaccine carrier systems. Here we show that α-Al(2)O(3) nanoparticles can act as an antigen carrier to reduce the amount of antigen required to activate T cells in vitro and in vivo. We found that α-Al(2)O(3) nanoparticles delivered antigens to autophagosomes in dendritic cells, which then presented the antigens to T cells through autophagy. Immunization of mice with α-Al(2)O(3) nanoparticles that are conjugated to either a model tumour antigen or autophagosomes derived from tumour cells resulted in tumour regression. These results suggest that α-Al(2)O(3) nanoparticles may be a promising adjuvant in the development of therapeutic cancer vaccines.     

A concentration dependent study of acoustic plate mode immunosensor response using antigen/antibody systems with different binding ability

Acoustic plate mode sensors have been used to monitor immunochemical reactions as a function of antigen concentration. In the studies, antibodies were covalently linked to the gold-coated sensing surface via mercaptoethanol, aminosilane, and glutaraldehyde. Two antigen/antibody model systems that differ in their ability to mutually bind one another have been used. For sensor operation at about 150 MHz, a detection limit of approximately 0.5 mug/ml was obtained in both cases. No significant difference between the two systems was found for the value of the binding constants. They amount to about 1.10(8) 1/mole and fall well into the range of binding constants reported for homogeneous immunoassays. A comparison of the sensor response obtained for the two model systems shows that about 70% of the immobilized antibodies are active.     

大菱鲆疾病早期快速检测方法——胶体金免疫层析试纸的研制与建立

使用成分单一的牛血清白蛋白(BSA)为模拟病原,以胶体金标记兔抗血清(即大菱鲆免疫球蛋白多抗)作为检测示踪物,并分别将BSA和葡萄球菌A蛋白印记到硝酸纤维素膜上制成检测线和对照线,通过一系列工艺创制与组装配套,首次成功制备了一套完整的大菱鲆抗体快速检测试纸。采用大菱鲆抗BSA血清作为阳性样本,以健康大菱鲆血清作为阴性样本,用以检验试纸的性能,并与酶联免疫吸附实验(ELISA)法检测结果相比较。结果表明:本试纸检测抗体的特异性与敏感性均很高,与ELISA方法相当,而且使用方便,不需专业技能和额外的试剂与辅助仪器设备,5 min内即可用裸眼获得观察结果,很适合于基层生产操作及户外调研使用。以该实验为基础建立起来的抗体检测试纸,亦可推广应用于其他病害抗体的检测,可为鱼类疾病早期发生提供简易、快捷和操作性强的诊断方法。     

Self-directed and self-oriented immobilization of antibody by protein G-DNA conjugate

A versatile biolinker for efficient antibody immobilization was prepared by site-specific coupling of protein G to DNA oligonucleotide. This protein G-DNA conjugate ensures the controlled immobilization of an antibody to the intended area on the surface of bioassay chips or particles, while maintaining the activity and orientation of the bound antibody. Streptococcus protein G tagged with a cysteine residue at the N-terminus was chemically linked to amine-modified, single-stranded DNA. SPR analysis indicated that the protein G-DNA conjugates sequence-specifically bind to complementary surface-bound DNA probes. More importantly, the resulting protein G, which is hybridized onto the DNA surface, possesses a greater antibody/antigen binding ability than even properly oriented protein G linked on the chip surface by chemical bonding. Antibody targeting on glass slides could also be achieved by using this linker system without modifying or spotting antibodies. Moreover, the protein G-DNA conjugate provided a simple but effective method to label DNA-functionalized gold nanoparticles with target antibodies. The DNA-linked protein G construct introduced in this study offers a useful strategy to manage antibody immobilization in many immunoassay systems.