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1.
从南海海绵Dysidea avara中分离筛选得到一株产弱碱性脲酶菌株Bacillus atrophaeus C89,为探究其在海洋环境重金属离子检测中的应用,在人工海水体系中研究Hg~(2+)、Cu~(2+)、Pb~(2+)和Cd~(2+)四种重金属离子对脲酶活性的影响。研究发现四种重金属离子的浓度与脲酶抑制率呈显著的正相关性,当仅有单一重金属离子存在时,各重金属离子对脲酶活性的抑制效应顺次为Hg~(2+)Cu~(2+)Cd~(2+)Pb~(2+);当有多种重金属离子同时存在时,联合抑制效应与抑制率较高的重金属离子接近,且不同组合存在不同的协同或拮抗效应,Hg~(2+)、Cu~(2+)、Cd~(2+)呈现协同效应,而Pb~(2+)表现出一定的拮抗效应。与刀豆脲酶对比发现,海洋微生物脲酶在p H和盐度等方面具有更高的耐受性,更适用于海洋环境重金属离子的检测。  相似文献   

2.
免疫测定法检测重金属技术的进展   总被引:1,自引:0,他引:1  
作为检测重金属的一种新型方法,免疫测定法有其特点和优势。免疫测定法的技术构成包括重金属单克隆抗体的获得、单克隆抗体灵敏性的提高和免疫检测,其中重金属单克隆抗体的获得是基础。我们对该方法进行了简要介绍,并就其所面临的一些问题和发展趋势进行了探讨。  相似文献   

3.
重金属残留超标是重大食品安全问题之一。为建立一种快速简单的Cd~(2+)和Pb~(2+)重金属离子检测法,以磁性Fe_3O_4纳米粒子(MNPs)的过氧化物酶活性作为传感信号输出工具,以核酸适配体的特异性识别功能作为靶标识别元件,建立了一个比色传感器。通过可行性分析和条件优化确定检测方法,再通过灵敏度和特异性实验对传感器性能进行评估。在最优条件下,Cd~(2+)和Pb~(2+)的最低检测线分别为0.108μmol/L和0.117μmol/L。该检测方法检测成本低、检测过程快速简单、检测结果肉眼可见,凭借手持式光度计即可实现定量分析。此外,该方法受其他离子干扰相对较小,特异性较强。该传感器的建立将使重金属的现场实时快速检测成为可能,为我国食品安全检测提供了有力保障。  相似文献   

4.
重金属离子对高等植物光合膜结构与功能的影响   总被引:60,自引:0,他引:60  
由于现代工业的迅速发展,重金属离子对土壤和水质的污染日趋严重,它们作为一种逆境因子胁迫着植物的各种生理过程,而光合作用比呼吸作用对重金属离子具有更高的敏感性。近年来已报道了一些关于重金属离子对高等植物光合膜结构和功能发生影响的工作。本文现对这方面的研究近况作一简要叙述,并着重讨论重金属离子对光系统Ⅱ的抑制作用。  相似文献   

5.
重金属残留的快速检测   总被引:7,自引:0,他引:7  
环境及农畜产品中的重金属残留已对人类安全构成严重威胁,急需快速、高效的重金属残留检测方法。该文总结了国内外有关采用生物、化学传感器、免疫学方法等快速检测环境及农畜产品中重金属残留的研究进展,并对其发展前景及市场化作了预测及展望。  相似文献   

6.
重金属对芝麻种子萌发及幼苗生长的影响   总被引:2,自引:0,他引:2  
采用培养皿滤纸发芽法研究镉(Cd2+)、镍(Ni2+)、铬(Cr6+)、汞(Hg2+)、铅(Pb2+)等5种重金属离子对芝麻Sesamum indicum种子萌发及幼苗生长的影响。结果表明,重金属离子胁迫对芝麻种子露白萌发影响较小,对胚根生长影响最大;随着重金属离子浓度增加,芝麻幼苗芽长、根长和鲜重减小;在Cd2+、Ni2+和Cr6+浓度20 mg·L-1,Hg2+浓度50 mg·L-1,Pb2+浓度100 mg·L-1时,芝麻种子萌发后胚根基本不生长,难以成苗,这些浓度可视为对应重金属离子对芝麻种子萌发毒害的致死浓度。综合比较,5种重金属离子对芝麻种子萌发及芽期幼苗生长的毒害程度依次为Ni2+Cr6+Cd2+Hg2+Pb2+。  相似文献   

7.
用硫酸盐还原菌处理重金属废水的研究   总被引:22,自引:0,他引:22  
介绍了用硫酸盐还原菌处理重金属废水的几种主要方法和原理。硫酸盐还原菌处理含重金属废水主要是通过将可溶性的重金属离子转化成不溶性的金属硫化物、氢氧化物、碳酸盐的方式 ,或直接通过以菌体对重金属离子的吸附完成的。目前研究用硫酸盐还原菌处理重金属废水的主要方法有分批沉淀工艺、吸附处理工艺、化学法和硫酸盐还原菌的混合工艺、全混合处理工艺及硫酸盐还原菌的厌氧上流式污泥床和流化床工艺 ,并对其主要的工艺指标进行了比较。  相似文献   

8.
抗重金属汞离子抗体的制备及鉴定   总被引:1,自引:0,他引:1       下载免费PDF全文
汞、镉、铅等重金属引起的环境污染已在世界范围内造成危害。快速、廉价地监测生境中重金属是减小其对人类及动物危害的先决条件。传统检测方法无法满足高通量的现场检测,建立更快速、更经济的免疫分析法检测汞离子是生产及经济发展的需要。本研究中,报道了汞特异性单克隆抗体的制备与筛选方法和结果。因Hg2+太小以至于不能引起免疫反应,所以用螯合剂(二乙烯三胺五乙酸,DTPA)将金属离子与载体蛋白(匙孔血蓝蛋白,KLH)连接起来。成功合成、鉴定汞复合物抗原后,免疫BALB/c小鼠,通过细胞融合获得了稳定分泌抗体的杂交瘤细胞。用极限稀释法亚克隆,通过ELISA筛选,获得了2株稳定分泌抗汞离子抗体的细胞株(H2H5,H1H8)。小鼠腹腔注射1×107H2H5、H1H8细胞株制备腹水,腹水抗体效价都在1∶51200以上。经鉴定两株杂交瘤均为IgG1亚类,轻链为kappa型且分泌抗体稳定性较好。实验结果为汞离子残留免疫学检测方法的建立提供了技术基础,对提高风险评估工作的效率和质量,保障食品安全有重要现实意义。  相似文献   

9.
高等植物重金属耐性与超积累特性及其分子机理研究   总被引:50,自引:0,他引:50       下载免费PDF全文
 由于重金属污染日益严重,重金属在土壤-植物系统中的行为引起了人们的高度重视。高等植物对重金属的耐性与积累性,已经成为污染生态学研究的热点。近年来,由于分子生态学等学科的发展,有关植物对重金属的解毒和耐性机理、重金属离子富集机制的研究取得了较大进展。高等植物对重金属的耐性和积累在种间和基因型之间存在很大差异。根系是重金属等土壤污染物进入植物的门户。根系分泌物改变重金属的生物有效性和毒性,并在植物吸收重金属的过程中发挥重要作用。土壤中的大部分重金属离子都是通过金属转运蛋白进入根细胞,并在植物体内进一步转运至液泡贮存。在重金属胁迫条件下植物螯合肽(PC)的合成是植物对胁迫的一种适应性反应。耐性基因型合成较多的PC,谷胱甘肽(GSH)是合成PC的前体,重金属与PC螯合并转移至液泡中贮存,从而达到解毒效果。金属硫蛋白(MTs)与PC一样,可以与重金属离子螯合,从而降低重金属离子的毒性。该文从分子水平上论述了根系分泌物、金属转运蛋白、MTs、PC、GSH在重金属耐性及超积累性中的作用,评述了近10年来这方面的研究进展,并在此基础上提出存在的问题和今后研究的重点。  相似文献   

10.
食品镉离子污染免疫检测技术的研究与应用   总被引:1,自引:0,他引:1  
传统的理化分析方法在食品Cd2+污染检测方面发挥了重要的作用,但均存在一定的缺陷,急需建立快速、高效的检测方法.食品Cd2+污染免疫检测是一种新型的分析方法,与理化分析方法相比,具有省时省力、费用低廉、便于携带、操作简便等优点,已成为重要的检测手段.详细综述了目前国内外在食品Cd2+污染免疫检测方面的研究方法和成果,主要包括Cd2+人工抗原的制备与鉴定、Cd2+单克隆抗体的制备及免疫学特性影响因素分析和Cd2+免疫检测方法的建立及应用.在总结这些研究的基础上,展望了食品Cd2+污染免疫检测技术的发展方向.  相似文献   

11.
As increasing incidences in the occurrence of cylindrospermopsin (CYN) appear, in addition to further research on its toxicological nature, improved rapid methods to detect this toxin are required. Antibody based assays are renowned for their ability to provide rapid, portable, simple to use tests. As yet however there are no publications outlining how an antibody to CYN can be produced. A range of chemical approaches was investigated to synthesise CYN immunogens for antibody production but failed to generate a response. Finally, a modified Mannich reaction for immunogen synthesis was employed to couple the toxin to two carrier proteins. Both protein conjugates were successfully used to raise both polyclonal and monoclonal antibodies of high sensitivity to CYN. These antibodies were characterised employing competitive indirect ELISA and an optical biosensor assay. By ELISA the sensitivity achieved ranged from 27 to 131 pg/mL and by SPR 4.4 to 11.1 ng/mL thus demonstrating that the selection of immunoassay platform is important for the detection level required by the end user for their application. Low cross-reactivity to the much less toxic metabolite deoxyCYN was observed. This is the first reported production of antibodies to this toxin.  相似文献   

12.
Evidence has accumulated that herbicides in the environment present a significant health hazard to the population. Therefore, the levels of heavily used substances such as atrazine and simazine and their metabolites need to be regularly assessed. The objective was to develop a rapid and simple tube ELISA procedure suitable for use in field studies and non-specialized laboratories. The antisera used were polyclonal antibodies raised in sheep against atrazine or simazine amido caproic acid conjugated to bovine serum albumin. The antibodies were first used to construct a two-step competitive ELISA procedure in 96-well microtitre plates. The 96-well format was then adapted to a coated-tube enzyme immunoassay, by immobilization of hapten-gelatine conjugates on polystyrene tubes. This enabled the colour to be read using a basic spectrophotometer. Soil samples were collected from agricultural and non-agricultural sites in Poland. Atrazine and simazine were extracted by liquid extraction from soil and assayed by tube ELISA. In addition, the samples were extracted by solid-phase extraction before analysis by HPLC. The immunoassays and chemical analysis were carried out by different individuals who were unaware of each other's results, which were then compared at the end of the study. Correlation of the two methods was excellent, with R=98.7 and 81.3 for atrazine and simazine, respectively. The immunoassay yielded the same order of results without having to perform solid-phase extraction before analysis. The study has demonstrated that the simple antigen-coated tube assay provides a cost-effective and valuable screening test. Comparison with the more elaborate, heavily labour-intensive HPLC analysis demonstrated that the results obtained by the simpler enzyme-immunoassay tests were within the same order.  相似文献   

13.
Evidence has accumulated that herbicides in the environment present a significant health hazard to the population. Therefore, the levels of heavily used substances such as atrazine and simazine and their metabolites need to be regularly assessed. The objective was to develop a rapid and simple tube ELISA procedure suitable for use in field studies and non-specialized laboratories. The antisera used were polyclonal antibodies raised in sheep against atrazine or simazine amido caproic acid conjugated to bovine serum albumin. The antibodies were first used to construct a two-step competitive ELISA procedure in 96-well microtitre plates. The 96-well format was then adapted to a coated-tube enzyme immunoassay, by immobilization of hapten-gelatine conjugates on polystyrene tubes. This enabled the colour to be read using a basic spectrophotometer. Soil samples were collected from agricultural and non-agricultural sites in Poland. Atrazine and simazine were extracted by liquid extraction from soil and assayed by tube ELISA. In addition, the samples were extracted by solid-phase extraction before analysis by HPLC. The immunoassays and chemical analysis were carried out by different individuals who were unaware of each other's results, which were then compared at the end of the study. Correlation of the two methods was excellent, with R=98.7 and 81.3 for atrazine and simazine, respectively. The immunoassay yielded the same order of results without having to perform solid-phase extraction before analysis. The study has demonstrated that the simple antigen-coated tube assay provides a cost-effective and valuable screening test. Comparison with the more elaborate, heavily labour-intensive HPLC analysis demonstrated that the results obtained by the simpler enzyme-immunoassay tests were within the same order.  相似文献   

14.
Prior observations that questioned the validity of kinetic exclusion assays were based on the mistaken assumption that the assays quantified the fraction of those antibody molecules that had unoccupied binding sites. Instead, the standard KinExA assay quantifies the fraction of total antibody binding sites that are unoccupied, regardless of the number of unoccupied sites on each antibody molecule. Although the standard KinExA analysis assumes that there is only a small probability of antibody-site capture by the affinity matrix, the results of numerical simulations demonstrate the reliability of dissociation constants obtained by the standard KinExA analysis for capture probabilities as high as 30%. This finding further strengthens the potential of kinetic exclusion assays as the procedure of choice for the rapid and accurate characterization of immunochemical reactions that forms part of screening processes in the search for therapeutic antibodies.  相似文献   

15.
Antibody-based sensors for heavy metal ions   总被引:13,自引:0,他引:13  
Competitive immunoassays for Cd(II), Co(II), Pb(II) and U(VI) were developed using identical reagents in two different assay formats, a competitive microwell format and an immunosensor format with the KinExA™ 3000. Four different monoclonal antibodies specific for complexes of EDTA–Cd(II), DTPA–Co(II), 2,9-dicarboxyl-1,10-phenanthroline–U(VI), or cyclohexyl–DTPA–Pb(II) were incubated with the appropriate soluble metal–chelate complex. In the microwell assay format, the immobilized version of the metal–chelate complex was present simultaneously in the assay mixture. In the KinExA format, the antibody was allowed to pre-equilibrate with the soluble metal-chelate complex, then the incubation mixture was rapidly passed through a microcolumn containing the immobilized metal-chelate complex. In all four assays, the KinExA format yielded an assay with 10–1000-fold greater sensitivity. The enhanced sensitivity of the KinExA format is most likely due to the differences in the affinity of the monoclonal antibodies for the soluble versus the immobilized metal–chelate complex. The KinExA 3000 instrument and the Cd(II)-specific antibody were used to construct a prototype assay that could correctly assess the concentration of cadmium spiked into a groundwater sample. Mean analytical recovery of added Cd(II) was 114.25±11.37%. The precision of the assay was satisfactory; coefficients of variation were 0.81–7.77% and 3.62–14.16% for within run and between run precision, respectively.  相似文献   

16.
江年  茆灿泉 《生物信息学》2009,7(4):284-287,291
金属离子与金属结合肽(蛋白)的相互作用与应用研究,一直是生物无机化学的重点和热点,也是分子间相互作用研究领域的难点。本研究利用ClustalX、BLAST等生物信息技术与方法对大量已知的重金属结合肽进行分析与数据挖掘。确定筛选获得的重金属结合肽常富含His,无Cys,无金属结合肽模式序列,进化不保守;部分氨基酸序列结构(如六肽)可在蛋白数据库中找到相似序列。序列特征主要为Zn^2+相关的转录因子。本研究为重金属结合蛋白-重金属离子的相互作用分析简化为重金属结合肽-重金属离子的结构模拟与分析提供了重要的理论基础和研究手段。  相似文献   

17.
The quantitation of low-molecular-weight haptens has been difficult with conventional sandwich immunoassays due to their small size. Many researchers have attempted to develop sandwich assays for haptens due to the significant advantages of the sandwich format over competitive assays including greater dynamic range, ease of automation, and sensitivity. Here we apply the open-sandwich ELISA (OS-ELISA), an immunoassay based on antigen-dependent stabilization of antibody variable regions (V(H) and V(L) domains), to hapten quantitation. Two fusion proteins, the high-affinity mutant V(H) domain from anti-4-hydroxy-3-nitrophenacetyl (NP) antibody B1-8 tethered with Escherichia coli alkaline phosphatase (V(H)(W33L)-PhoA) and the V(L) domain from the same antibody tethered with Streptococcus sp. protein G, were made. These fusion proteins when added together achieved Fv reassociation consequent to the addition of NP. Signal was generated in a direct relationship to the NP concentration with better sensitivity compared with competitive immunoassay, demonstrating this assay to be a quick noncompetitive alternative to the conventional assays for small compounds, such as environmental pollutants, drugs of abuse, and therapeutic drugs. With our previous demonstration that the OS-ELISA works well with large proteins, the OS-ELISA becomes the first practical immunoassay approach capable of quantifying any molecule regardless of their size.  相似文献   

18.
We have investigated the complex formation between an immobilized monoclonal antibody and antigens that differ in size about 50-fold. As a model system, we used an iodinated progesterone derivative and a progesterone-horseradish peroxidase conjugate as tracer and a monoclonal antibody as binding protein. The antibody was immobilized by four different methods: physical adsorption, chemical binding, and binding via protein G in the absence or presence of a protective protein (gelatin). These investigations have shown that the performance of competitive immunoassays is determined by a combination of factors: (a) the relative size of the analyte and the tracer, (b) the antibody density on the solid matrix, (c) the method of immobilization of the antibody, and (d) the binding constants between antibody-analyte and antibody-tracer. All of these interactions have to be considered in designing an optimal immunoassay. The smaller antigen can form a 3- to 35-fold higher maximal complex density than the larger antigen. Dose-response curves are less affected by the size of the tracer than by the binding constant with the antibody. A large enzyme tracer with a relatively low binding constant can, therefore, provide a more sensitive assay. On the other hand, the increase in complex density achieved with a smaller tracer yields a higher signal that in turn can provide a better signal-to-noise ratio in highly sensitive competitive solid-phase immunoassays. We have suggested a model for antibody immobilization that accounts for the interdependence of tracer size, complex formation, and antibody density. The methods described can be used to design and optimize immunoassays of predefined performance characteristics. The results are particularly useful for converting radioimmunoassays to enzyme immunoassays.  相似文献   

19.
The objectives of this study were to develop an enzyme immunoassay for metallothioneins in human urine using a polyclonal antiserum and to demonstrate a possible relationship between the level of this biomarker and heavy metal exposure. The antiserum was raised in sheep against horse metallothionein conjugated to carboxylated bovine serum albumin. The antibody was used to construct a two-step competitive ELISA procedure. Human urine was treated with activated charcoal powder to remove traces of metallothioneins and known amounts of pure metallothioneins were added to provide standards for a standard curve. Metallothionein levels were measured in two groups of children living in areas of mild and high environmental pollution due mainly to heavy metals. A comparison was made between the biomarker levels and the levels of cadmium and lead in urine samples in the two groups. A group of children from a non-polluted area acted as controls. The results show that the detected levels of metallothioneins appear to correspond to levels of the two heavy metals studied and that there was an apparent relationship to the environmental exposure. Thus according to results of this study the increase in the metallothionein excretion seems to provide an indication of previous of exposure to metals. The ELISA procedure is sensitive and robust and can be used to screen large numbers of samples and is more rapid than the physical procedures currently used for analysis of these proteins. The assay can therefore be used as an additional tool for screening at-risk populations where either environmental or occupational exposure to divalent heavy metals is suspected.  相似文献   

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