FTB多克隆抗体的研制

为建立ＦＴＢ的间接酶联免疫吸附方法，用自行制备的ＦＴＢＳ－载体蛋白结合物ＦＴＢＳ－ＢＳＡ和ＦＴＢＳ－ＨＳＡ作为免疫原，按常规免疫程序分别免疫新西兰大耳白家兔各２只，４～１０周后获得２种抗ＦＴＢ的多克隆抗体，ＰｃＡｂ－ＦＴＢ１（以ＦＴＢＳ－ＢＳＡ为免疫原）和ＰｃＡｂ－ＦＴＢ２（以ＦＴＢＳ－ＨＳＡ为免疫原）。以ＦＴＢＳ－ＩｇＧ作为标识抗原包被酶标板，建立了测定ＦＴＢ的间接酶联免疫吸附试验（ＩＣ－ＥＬＩＳＡ）方法，这２株多克隆抗体与８种真菌毒素的交叉反应均较小。     

酱油中黄曲霉素素B1的酶联免疫检测

本研究将间接竞争ＥＬＩＳＡ用于酱油中ＡＦＢ１的检测。和ＴＬＣ法相比，该法有灵敏、快速、特异等优点。研究中首先对样品毒素的几种提取方法进行了比较，结果表明甲醇－－二甲基甲酰胺提取法是最好的毒素提取方法，它的样品提取液对ＥＬＩＳＡ标准曲线无影响而且毒素的提取时间较短。在此基础上进行了ＡＦＢ１的回收试验，ＡＦＢ１的浓度为３－８０μｇ／ｋｇ时，ＡＦＢ１的ＥＬＩＳＡ回收效果好，回收变异系数都小于１０％。进一     

美国几项最新非织造布专利

美国几项最新非织造布专利·ＵＳＰ５３５５５６５１９９４．１０．１８（ＦｒｅｎｄｅｎｂｅｒｇＳｐｕｎｗｅｂ）采用２ＤＰＥＴ／ＰＡ、ＰＥＴ／ＰＰ、ＰＡ／ＰＰ并列型复合纤维网作水刺法非织造布，布重２０～２５０９／ｍ２。水刺时，复合纤维分裂成２根１Ｄ纤维。车...     

第八届北京木工机械展主要厂商及展品简介

公司名称产品及业务（续） ＩＴＡＬＹ意大利Ａ ＣＯＳＴＡ Ｓ．Ｐ．Ａ．封边机、横切机、木地板生 产线ＡＬＦＡＭＡＣＣＨＩＮＥ Ｄ ＲＡＦ－框架装配机、双横切锯ＦＯＮＩ ＧＩＵＳＥＰＰＥＡＮＧＥＬＯ ＣＲＥＭＯＮＡ原木制备设备Ｓ．Ｐ．Ａ．ＡＲＴＵＲＯ ＳＡＬＩＣＥ Ｓ．Ｐ、Ａ．家俱自动关闭铰链ＡＵＴＥＣＯ Ｓ．Ｒ．Ｌ．多轴四面铣刨机ＢＡＬＥＳＴＲＩＮＩ ＲＥＮＺＯ实木餐椅生产机械Ｓ．Ｐ．Ａ．ＢＩＥＳＳＥ ＧＲＯＵＰ木工机械ＢＭＡ ＡＢＲＡＳＩＶＥＳ砂纸ＢＲＥＶＥＴＴＩＭＡ双锯斜角切割机、固定、涂 胶、基砖材料接…     

酱油中黄曲霉毒素B_1的酶联免疫检测

本研究将间接竞争ＥＬＩＳＡ用于酱油中ＡＦＢ１的检测。和ＴＬＣ法相比，该法有灵敏、快速、特异等优点。研究中首先对样品毒素的几种提取方法进行了比较，结果表明甲醇—二甲基甲酰胺提取法是最好的毒素提取方法。它的样品提取液对ＥＬＩＳＡ标准曲线无影响而且毒素的提取时间较短。在此基础上进行了ＡＦＢ１的回收试验，ＡＦＢ１的浓度为３—８０μｇ／ｋｇ时，ＡＦＢ１的ＥＬＩＳＡ回收效果好，回收变异系数都小于１０％。进一步对从市场购买采集的酱油样品中的ＡＦＢ１以ＥＬＩＳＡ进行了分析。     

复合甜味剂(二)

图２　Ｈｉｓ及ＢｌｅｎｄＨｉｓ苦味／后味特性显著特性组成区域分布图１－Ｓｕｒｏｓｅ　２－ＡＰＭ　３－ＴＧＳ　４－ＣＹＣ　５－ＳＡＣ　６－ＡＣＫ　７－ＡＣＫ／ＡＰＫ　８－ＡＣＫ／ＮＨＤＣ　９－ＡＣＫ／ＴＧＳ　１０－ＡＣＫ／ＣＹＣ　１１－ＣＹＣ／ＳＡＣ　１２－ＡＣＫ／ＡＰＭ／ＣＹＣ　１３－ＡＣＫ／ＡＰＭ／ＳＡＣ　１４－ＡＣＫ／ＡＰＭ／ＳＡＣ／ＣＹＣ　　另从表２可见,在苦味、ｂｉｔｔｅｒ－ＡＴ、ｂｉｔｔｅｒ－ＡＴ２、ｏｆｆ－ｆｌａｖｏｒ－ＡＴ这些特性上,所有的Ｈｉｓ、ＢｌｅｎｄＨｉｓ与蔗糖相互间无区别…     

烟草上马铃薯Y病毒的提纯与检测

本文是关于制备马铃薯Ｙ病毒抗血清及ＥＬＩＳＡ检测技术的实验结果。采用垫层差速离心、蔗糖密度梯度离心提纯ＰＶＹ,纯化的ＰＶＹＯＤ２６０／ＯＤ２８０为１．５６,将提纯的病毒制品免疫家兔,制备抗血清,经微量沉淀及环状沉淀试验,用提纯病毒测定抗血清的效价均为１／２０４８,并对制备的ＰＶＹ—ＩｇＧ作灵敏度及特异性测定。     

制版新概念

丹麦宝宝禄德福公司生产的新概念"版神"ＩＭＡＭＧＥＭＡＫＥＲ系列是该公司推出的高效计算机直接胶片／印版输出系统。该系列输出机有Ｂ１／Ｂ２ＣＴＦ胶片输出机和Ｂ１／Ｂ２ＣＴＰ印版输出机四种基本型号，Ｂ２幅百可现场升级到Ｂ１幅面，输出胶片的ＣＴＰ可现场升级到输出印版的ＣＴＰ，并最终可升级到半自动或全自动的胶片／印版输出两用机。详情请洽：０１０－６４９１７２５０．６４９１７２８１制版新概念     

北京赛恩思科贸有限公司食品检测试剂盒

β -兴奋剂 (盐酸克伦特罗 )ＥＬＩＳＡ检测试剂盒 ,测定简单 ,方便 ,快速 ,灵敏度可达到 0 1ｐｐｂ ,适合各级卫生防疫部门使用。德国拜发公司ｒ Ｂｉｏ ｐｈａｒｍ将在 2 0 0 1年第三季度推出疯牛病ＥＬＩＳＡ检测试剂盒。德国拜发公司ｒ Ｂｉｏｐｈａｒｍ检测盒霉菌毒素ＥＬＩＳＡ检测试剂盒黄曲霉毒素Ｂ1 　ＡｆｌａｔｏｘｉｎＢ1黄曲霉毒素总量　ＡｆｌａｔｏｘｉｎＴｏｔａｌ黄曲霉毒素的快速检测ＦＡＳＴＡｆｌａｔｏｘｉｎ黄曲霉毒素免疫亲和层析柱ＡｆｌａｔｏｘｉｎｃｏｌｕｍｎＴ -2毒素的快速检测ＦＡＳＴＴ -2Ｔｏｘｉｎ赭曲…     

绵羊服装革工艺

ＴｅｃｈｎｉｑｕｅｏｆＳｈｅｅｐｓｋｉｎＧａｒｍｅｎｔＬｅａｔｈｅｒ特约栏目主持：肖传斌（德国司马化学有限公司上海办事处）垂询电话：（０２１）－６４４００６８２）１原材料材料：国产绵羊蓝湿皮厚度：０．８～１．０ｍｍ用料参照削匀皮重量　２水洗２００％　水,３５℃１．０％　ＳＵＰＲＡＬＡＮ　８０（非离子型脱脂剂）０．１%　甲酸４０分钟ｐＨ４．０排水３铬复鞣／中和１００％　水,４０℃１．５％　ＰＥＬＧＲＡＳＳＯＬ　ＳＦ（耐电解质合成加脂剂）１,５％　ＰＲＯＶＯＬ　ＢＡ１０分钟２．０％　ＮＯＶＡＬＴＡＮ…     

弧菌外膜蛋白OmpK单克隆抗体的制备及其特性

制备弧菌外膜蛋白K(outer membrane protein K,Omp K)单克隆抗体并对其特性进行研究。以原核表达系统表达野生株Vibrio parahaemolyticus B的Omp K免疫Balb/c小鼠,取免疫小鼠脾细胞与肿瘤细胞SP2/0进行细胞融合,采用有限稀释法和间接酶联免疫吸附测定法筛选出能够稳定分泌单克隆抗体的杂交瘤细胞株,小鼠体内诱导制备腹水,用饱和硫酸铵沉淀法和亲和层析柱纯化抗体。最终获得能稳定分泌抗Omp K的两株单克隆抗体杂交瘤细胞株Omp K-Mab-4B7、Omp K-Mab-3C5,2株杂交瘤细胞系所分泌的Mab亚类均为Ig G1,效价达1∶128 000,抗体3C5、4B7的敏感度IC50分别为2.5、5.0μg/m L。Western blotting结果显示单克隆抗体可以与本实验室12株副溶血性弧菌(V.parahaemolyticus A、B、C、D、E、F、G、H、I、J、ATCC17802、ATCC33847)的外膜蛋白有不同程度的结合反应,与4株溶藻弧菌中的3株(V.alginolyticus A、B、C)外膜蛋白能够较好的结合,与1株鳗弧菌(V.anguillarumMVM)外膜蛋白也有轻微的结合反应,实验制备的单克隆抗体可用于弧菌Omp K的基础研究和快速检测。     

芝麻过敏原油质蛋白O leosins单克隆抗体的制备与鉴定

利用细胞融合技术制备小鼠抗油质蛋白杂交瘤细胞,通过间接ELISA方法筛选稳定分泌特异性抗体的杂交瘤细胞株。获得10株杂交瘤细胞,分别命名为1B2、2D8、2F3、3A4、3D7、3F8、4A10、4F12、6A12、6E7,利用Ig亚类鉴定试剂盒鉴定各单克隆抗体的Ig亚型,除1B2、3D7、3F8为IgG1类外其余均为IgG2a类型。将杂交瘤细胞株注射入小鼠腹腔获取腹水,应用Protein A亲和层析法对腹水进行纯化。单抗效价有9株在2.0×105以上。ELISA和Western blotting分析表明这些单抗均能特异性识别芝麻过敏原油质蛋白。     

免疫亲和柱净化-光化学柱后衍生高效液相色谱 荧光法测定粮食中黄曲霉毒素

建立了粮食中黄曲霉毒素B1、B2、G1、G2的免疫亲和柱净化-光化学柱后衍生高效液相色谱荧光检测法。样品经甲醇-水提取,免疫亲和柱净化,高效液相色谱分离,光化学柱后衍生,荧光检测器测定。结果表明,黄曲霉毒素B1、B2、G1、G2的检出限分别为0.50、0.25、0.50、0.25μg/kg,回收率为67.2%～91.7%,RSD小于10%。该方法快速、准确、灵敏度高、重现性好,能满足我国对粮食中黄曲霉毒素限量的检测要求。     

Influence of a cocoa-enriched diet on specific immune response in ovalbumin-sensitized rats

Previous studies in young rats have reported the impact of 3 weeks of high cocoa intake on healthy immune status. The present article describes the effects of a longer-term cocoa-enriched diet (9 weeks) on the specific immune response to ovalbumin (OVA) in adult Wistar rats. At 4 weeks after immunization, control rats produced anti-OVA antibodies, which, according their amount and isotype, were arranged as follows: IgG1 > IgG2a > IgM > IgG2b > IgG2c. Both cocoa diets studied (4% and 10%) down-modulated OVA-specific antibody levels of IgG1 (main subclass associated with the Th2 immune response in rats), IgG2a, IgG2c and IgM isotypes. Conversely, cocoa-fed rats presented equal or higher levels of anti-OVA IgG2b antibodies (subclass linked to the Th1 response). Spleen and lymph node cells from OVA-immunized control and cocoa-fed animals proliferated similarly under OVA stimulation. However, spleen cells from cocoa-fed animals showed decreased interleukin-4 secretion (main Th2 cytokine), and lymph node cells from the same rats displayed higher interferon-gamma secretion (main Th1 cytokine). These changes were accompanied by a reduction in the number of anti-OVA IgG-secreting cells in spleen. In conclusion, cocoa diets induced attenuation of antibody synthesis that may be attributable to specific down-regulation of the Th2 immune response.     

阪崎克罗诺杆菌单克隆抗体的制备及其间接竞争-ELISA检测方法

以阪崎克罗诺杆菌标准菌株ATCC 29004免疫BALB/c小鼠,采用杂交瘤细胞技术,经过两次细胞融合共制备出7株分泌单克隆抗体的杂交瘤细胞株(1E9、2A7、2D10、3E5、5B10、6E3、12A2)。其中2A7、2D10、3E5、12A2的效价达到了1∶256 000,6E3的效价为1∶128 000,5B10的效价为1∶64 000,1E9的效价为1∶32 000。亚型鉴定结果显示1E9的亚型为Ig G1,6E3的亚型为Ig G2b,其余抗体2A7、2D10、3E5、5B10、12A2的亚型均为IgG2a。对7株抗体进行特异性检测结果显示,2A7能与3株阪崎克罗诺杆菌(ATCC 29004、ATCC 29544、ATCC 12868)都结合,1E9能与其中两株(ATCC 29004、ATCC 12868)结合,且与其他11株类似菌和常见致病菌交叉反应结果显示7个抗体均有良好的特异性。用2A7建立的间接竞争酶联免疫吸附测定法对阪崎克罗诺杆菌的检测灵敏度可达到10~6 CFU/m L。     

Assessment of the opsonic activity of purified bovine sIgA following intramammary immunization of cows with Staphylococcus aureus

The phagocytosis of Staphylococcus aureus by bovine polymorphonuclear neutrophils (PMN) requires the presence of antibodies. Among the major isotypes of bovine antibodies, IgG2 and IgM are considered opsonic for bovine PMN. However, the role of purified bovine secretory IgA (sIgA) as an opsonin has not been assessed. In the present study, IgG2 were obtained from serum and sIgA, IgG1, and IgM were purified from the colostrums of three cows intramammarily immunized with heat-killed Staphylococcus aureus. The Ig preparations were assayed for specific antibodies, and the opsonic capacity of every isotype was investigated. Despite the presence of antibodies, we observed no distinct chemiluminescence response of PMN stimulated with sIgA- or IgG1-opsonized S. aureus, whereas IgM or IgG2 bound to bacteria induced a marked chemiluminescence response. Moreover, the counting of internalized bacteria per PMN after phagocytosis revealed a low uptake of S. aureus opsonized with sIgA or IgG1, in contrast to IgM or IgG2, which triggered efficient ingestion of bacteria. Priming of neutrophils by TNF-alpha, IFN-gamma, or C5adesArg did not promote an oxidative burst or uptake of sIgA-opsonized S. aureus to a greater extent than with IgG1-opsonized bacteria. Furthermore, analysis of uningested bacteria by flow cytometry after incubation with PMN showed a preferential uptake of IgM-opsonized S. aureus by PMN and only few sIgA-positive stained bacteria were PMN-associated. These experiments indicate that sIgA, like IgG1 and unlike IgM or IgG2, could not be considered as a major opsonin for phagocytosis of S. aureus by bovine blood PMN.     

高有机溶剂耐受性的苯并(a)芘单克隆抗体制备及其免疫反应特性研究

目的 制备适应油脂样品中苯并(a)芘[Benzo(a)pyrene,BaP]检测需求的抗BaP单克隆抗体。方法 利用杂交瘤单克隆抗体技术制备抗BaP的抗体,在抗体筛选过程中使用高浓度的有机溶剂(50%甲醇)来选择具有较高有机溶剂耐受能力的抗体,并利用间接竞争酶联免疫吸附法考察了抗体的免疫反应特性。结果 筛选获得一株对有机溶剂有较高耐受能力的、稳定分泌BaP单克隆抗体的细胞株2H10,该抗体为IgG1亚型,可耐受高浓度的甲醇、丙酮、二甲亚砜(Dimethyl sulfoxide,DMSO)和二甲基甲酰胺(Dimethylformamide,DMF)。60%甲醇和60% DMSO作为样品稀释液时,抗体对BaP的IC₅₀分别为81.4和24.5 ng/mL,以60% DMSO作为样品稀释液,可以显著降低BaP抗体对其它多环芳烃化合物的交叉反应率。结论 本研究获得高特异性的BaP人工抗原及高有机溶剂耐受性的抗BaP单克隆抗体。     

Serum immunoglobulin profiles of dairy cows chronically treated with recombinant bovine somatotropin

The objective of this study was to determine the effects of exogenous somatotropin on blood profiles of the major bovine Ig isotypes. Holsteins were treated with either 0, 10.3, or 20.6 mg recombinant bST/d. Subcutaneous injections were started between wk 4 and 5 of lactation and continued for 266 d. Blood samples were collected by coccygeal venipuncture throughout lactation for serum IgG, G1, G2, A, and M concentration determination. Cows treated with 10.3 mg of bST/d exhibited modest increases in mean blood concentrations of IgG (12.4%) and IgG2 (18.4%) relative to the other groups of cows. In addition, treated cows had 10.1% higher average concentration of IgA than controls. Somatotropin did not influence average lactational concentrations of IgG1 or IgM, although the group receiving 10.3 mg/d had higher late lactation concentrations of IgG1 than did the other two groups. However, treatment mean isotype concentrations were always within normal ranges, except for IgG2 from the 10.3 mg/d group, which were higher. The practical significance of elevated serum Ig concentrations observed in the present study is not known. However, the present data do show that bST administered at doses that increased milk yield had no apparent detrimental effect on humoral immunity as measured by blood concentrations of Ig.     

Rapid analysis of aflatoxins in raw peanuts,corn, buckwheat and red pepper by a new mini-column cleanup and HPLC using post-column photochemical derivatization system

A rapid and clean method for the analysis of aflatoxins (AFs) was developed by using a new column and post-column photochemical derivatization HPLC with fluorescence detection. The new cleanup column consisted of magnesia and basic alumina poured on the top of a commercial multi-functional mini-column. It was extremely effective for the cleanup of AFs from raw peanut, corn, buckwheat and red pepper. Fluorescent substances, which interfered with the analysis of AFs from corn, were completely absorbed at the top of the magnesia layer. Recoveries of AFs (B1, B2, G1, G2) added to raw peanuts, corn, buckwheat and red pepper were over 80% at two levels of fortification (higher level: 10, 3, 10, 3 ng/g, respectively, lower level: 1.0, 0.3, 1.0, 0.3 ng/g, respectively). Coefficients of variation were smaller than 12%, except the lower fortified level for red pepper. Limits of detection for AFs in raw peanuts, corn and buckwheat were 0.3 ng/g for B1 and G1, and 0.1 ng/g for B2 and G2. Those in red pepper were 0.5 ng/g for B1, B2, G1 and G2.     

PREVENTION OF CROSS-REACTIONS IN THE ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) FOR THE DETECTION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B IN CULTURE FILTRATES AND FOODS

Investigations were performed to avoid cross-reactions in the enzyme linked immunosorbent assay, using the sandwich technique, for detection of Staphylococcus aureus enterotoxin type B. Non-specific reactions can be caused by cross-reacting antigens and by Protein A, produced by S. aureus. The former reactions can be prevented by adsorption with culture filtrate of non-toxin type B producing strains. The latter reaction is caused by the binding of Protein A to the Fc fragments of the IgG antibodies. Interference by Protein A was completely eliminated by using the F(ab')₂ fragments of the IgG antibodies.
ELISA experiments in which these purified F(ab')₂ fragments were used resulted in a highly specific detection of entertoxin type B, both in culture filtrates and in foods.