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1.
重金属铅酶联免疫检测方法的研究   总被引:1,自引:0,他引:1  
制备铅离子多克隆抗体,并建立间接竞争ELISA检测方法。利用双功能螯合剂ITCBE将Pb2+与载体蛋白KLH和BSA偶联在一起,制备免疫抗原与检测抗原;通过免疫新西兰大耳白免成功制备铅多克隆抗体,建立间接竞争ELISA检测方法,并与ICP-MS检测结果进行比较。标准曲线IC50为3.48ng/mL,IC15为0.32ng/mL,与ICP-MS法检测结果的总符合率超过97%。  相似文献   

2.
沙丁胺醇单克隆抗体的制备及间接ELISA方法的建立   总被引:2,自引:0,他引:2  
制备抗沙丁胺醇单克隆抗体,鉴定其特异性并建立间接竞争ELISA检测方法.以合成的沙丁胺醇完全抗原免疫Balb/c小鼠,利用杂交瘤技术进行细胞融合,筛选克隆得到稳定分泌抗沙丁胺醇抗体的单克隆杂交瘤细胞,命名为4E4和3A2.经鉴定,两个抗体亚类都为IgG,,其腹水纯化后效价分别达到2.56 ×105和1.28 ×105.以效价高且特异性较好的4E4细胞株获得的抗体建立间接竞争ELISA方法,经条件优化后得到标准曲线,检测范围为4.76~84.99ng/mL,IC50为20.12ng/mL,检测限为3.25ng/mL,与克伦特罗交叉反应率为253.62%,与其它结构类似物没有明显交叉反应.本研究制备的单克隆抗体和建立的间接ELISA方法,可用于开发同时检测沙丁胺醇和克伦特罗的试剂盒.  相似文献   

3.
孙晶玮  胡章立 《现代食品科技》2013,29(11):2712-2716
设计并成功合成了三种孔雀石绿半抗原,所有半抗原均采用活化酯法分别与血匙兰蛋白(KLH)偶联制备成免疫抗原,与卵清蛋白(OVA)偶联制备成包被抗原。利用所制备的三种免疫原免疫新西兰大耳白兔都获得了高效价的抗孔雀石绿抗体,并将每一种抗体都与三种包被抗原进行组合配对,通过间接竞争酶联免疫吸附检测(ELISA)方法筛选出最佳的抗体-包被抗原组合,筛选出的抗体采用Sepharose FF-Protein A亲和层析柱纯化,采用间接ELISA法测定效价及鉴定特异性。经测定筛选出的抗体效价达到1:12000,IC50值为0.65 ng/mL,交叉反应表明该抗体有较好的特异性,与结构类似物结晶紫的交叉反应率为18.73%,与隐性孔雀石绿及隐性结晶紫的交叉反应率低于10%。本研究为建立快速检测食品中孔雀石绿残留的免疫分析方法奠定了基础。  相似文献   

4.
目的建立小麦球蛋白的ELISA检测技术,用于蛋白质掺假以及过敏原成分的快速检测。方法从麦胚粉中提取球蛋白,抗原免疫Balb/c小鼠进行4次免疫后,通过脾脏细胞杂交瘤技术及间接ELISA筛选制备单克隆抗体,同时制备兔抗小麦球蛋白的多克隆抗体。通过棋盘滴定法,初步确定单克隆抗体和多克隆抗体的最佳工作浓度,建立双抗夹心ELISA。结果通过免疫和杂交瘤技术获得了抗小麦球蛋白的单克隆抗体,纯化后抗体的效价均达到1:107,通过免疫兔制备的多克隆抗体经纯化后效价在1:2.4×105左右,所建立的双抗体夹心ELISA方法最低检测限为10 ng/m L,与其他物种的蛋白不发生交叉反应。结论本文建立的双抗体夹心ELISA方法具有良好的特异性和灵敏度,为建立乳品中小麦成分掺假及过敏原成分快速检测提供理论依据。  相似文献   

5.
合成具有阿维菌素特征结构的半抗原,采用活化酯法与载体蛋白BSA偶联制备人工抗原.通过免疫新西兰大白兔成功制备阿维菌素多克隆抗体,抗体效价达1∶20 000,用此抗体建立的阿维菌素间接竞争酶联免疫标准曲线IC50为1.92 ng/mL,IC15为0.13 ng/mL.  相似文献   

6.
氯霉素人工抗原的合成及多克隆抗体的制备   总被引:1,自引:0,他引:1  
采用重氮化法合成氯霉素人工抗原.通过紫外扫描、凝胶电泳、基质辅助激光解吸飞行时间质谱(MAL-DI-TOFMS)鉴定人工抗原.用该人工抗原免疫兔子,获得多抗血清.采用酶联免疫法测定抗体效价,并对ELISA条件进行初步优化.选择与氯霉素结构相似的4种药物进行交叉反应,计算交叉反应率.结果表明免疫抗原偶联率为1:22,最佳包被抗原质量浓度为0.25μg/mL,抗体效价128 000.优化后,通过间接竞争ELISA法建立标准曲线,IC50为0.26μg/mL,4种药物与抗体的交叉反应率均小于0.1%,特异性显著.  相似文献   

7.
采用优化的碳二亚胺法制备诺氟沙星(NOR)人工抗原,免疫新西兰大白兔制备多克隆抗体,建立间接竞争ELISA标准曲线。对NOR的线性检测范围为0.07~31.8ng/mL,半数抑制质量浓度(IC50)和最低检测限(LOD)分别为1.5ng/mL和0.04ng/mL。检测缓冲液中pH值和磷酸根离子浓度的最佳参数分别为7.4和10mmol/L;对甲醇和NaOH溶液(30mmol/L)的最佳调整体积分数分别为30%和10%。抗体具有广谱特异性,与环丙沙星(83.3%)、培氟沙星(78.9%)、依诺沙星(68.2%)、恩诺沙星(51.7%)和洛美沙星(41.7%)有较高的交叉反应率,可用于氟喹诺酮类药物(FQs)多残留检测方法研究。  相似文献   

8.
牛奶中三聚氰胺的ELISA检测研究   总被引:1,自引:0,他引:1  
戊二醛法将三聚氰胺(Melamine)与载体蛋白进行偶联制备免疫抗原Melamine-BSA及包被抗原Melamine-OVA.对人工合成抗原进行紫外扫描,其紫外扫描谱图与载体蛋白及三聚氰胺的图谱有明显的差异,初步显示载体蛋白与三聚氰胺偶联成功.进一步分析Melamine-BSA中Melamine与BSA的摩尔比,复合物中三聚氰胺与BSA的摩尔比为8.56 ∶ 1.用免疫抗原Melamine-BSA腹腔免疫BALB/c小白鼠,获得抗三聚氰胺的多克隆抗体,间接非竞争ELISA分析所得抗血清的效价达10000以上.间接竞争ELISA显示所制备的抗血清能有效识别三聚氰胺,与氰脲酸、三聚氰酸酰胺、三聚氰酸二酰胺无交叉反应.利用自制抗体建立牛奶中三聚氰胺的间接竞争ELISA检测方法,对奶样作0.1μg/mL,1μg/mL,10μg/mL,100μg/mL,500μg/mL和1000μg/mL六个加标浓度的回收率在70.30%113.59%.  相似文献   

9.
本文研究了环丙沙星(CPLX)羧基衍生物及其抗体的制备,并对该衍生物抗体的特异性进行了测定。将环丙沙星与3-溴丙酸反应,生成环丙沙星的羧基衍生物(CPLX-COOH),通过制备液相进行纯化,用红外光谱(IR)、电喷雾电离质谱(ESI-MS)与核磁共振法(NMR)进行鉴定;将环丙沙星衍生物分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)进行偶联,分别制备免疫抗原(CPLX-COOH-BSA)和包被抗原(CPLX-COOH-OVA),并用紫外光谱法进行鉴定。用免疫抗原免疫BALB/c小鼠,制备环丙沙星衍生物多克隆抗体。通过ELISA方阵滴定法,确定包被抗原的最佳包被浓度和抗体最佳稀释度,建立了CPLX-COOH间接竞争ELISA检测方法,其半抑制浓度IC50为228.56 ng/mL,最低检出限(LOD)为25.527 ng/mL,该抗体具有广谱特异性,可与多种氟喹诺酮类药物发生交叉反应,可进行氟喹诺酮类药物残留的多元检测。  相似文献   

10.
目的:合成铅离子完全抗原,并对其进行鉴定;用合成的完全抗原免疫Balb/c 小鼠制备多克隆抗体,鉴定效价和特异性。方法:用双功能螯合剂p-SCN-Bn-DTPA 将Pb2+ 与载体蛋白BSA 和OVA 偶联在一起,合成免疫抗原(Pb-DTPA-BSA)和检测抗原(Pb-DTPA-OVA),并通过SDS-PAGE、紫外扫描、电感耦合等离子体(ICP) 检测抗原中Pb2+ 含量,以及通过免疫Balb/c 小鼠,采集血清进行间接ELISA 和竞争ELISA 检测。结果:两种合成的抗原相对于各自的载体蛋白均出现了蛋白条带滞后,紫外吸收峰偏移等现象;免疫的4 号和5 号Balb/c 小鼠的抗血清效价达1:400000;与OVA 无交叉反应,ELISA 检测IC50 值为1.5ng/mL,与Fe3+ 有近5% 的交叉反应,与其他重金属离子的交叉反应均小于1%。结论:铅离子多克隆抗体制备成功。  相似文献   

11.
Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.  相似文献   

12.
An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.  相似文献   

13.
The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.  相似文献   

14.
The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.  相似文献   

15.
BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H2O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.  相似文献   

16.
17.
A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.  相似文献   

18.
19.
This paper describes the second part of a project undertaken to develop certified mussel reference materials for paralytic shellfish poisoning toxins. In the first part two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin and decarbamoyl-saxitoxin in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the certification exercise. Fifteen laboratories participated in this certification study and were asked to measure saxitoxin and decarbamoyl-saxitoxin in rehydrated lyophilized mussel material and in a saxitoxin-enriched mussel material. The participants were allowed to use a method of their choice but with an extraction procedure to be strictly followed. The study included extra experiments to verify the detection limits for both saxitoxin and decarbamoyl-saxitoxin. Most participants (13 of 15) were able to meet all the criteria set for the certification study. Results for saxitoxin.2HCl yielded a certified mass fraction of <0.07 mg/kg in the rehydrated lyophilized mussels. Results obtained for decarbamoyl-saxitoxin.2HCl yielded a certified mass fraction of 1.59+/-0.20 mg/kg. The results for saxitoxin.2HCl in enriched blank mussel yielded a certified mass fraction of 0.48 +/- 0.06 mg/kg. These certified reference materials for paralytic shellfish poisoning toxins in lyophilized mussel material are the first available for laboratories to test their method for accuracy and performance.  相似文献   

20.
<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of the  相似文献   

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