检索结果-内蒙古大学图书馆

中国临床康复 2006年第45期10卷 100-103页

作者：王美亮朱勇王九平金伯泉解放军第四军医大学免疫学教研室陕西省西安市710032 解放军第四军医大学唐都医院全军感染病诊疗中心陕西省西安市710038

目的:采用人工神经网络模型及在线软件BIMAS和SYFPEITHI共同预测汉滩病毒核蛋白HLA-A)02限制的CTL表位,并对CTL表位进行实验鉴定。方法:①选取2004-11解放军第四军医大学唐都医院住院的肾综合征出血热患者1例,汉滩病毒特异性IgM抗体为... 详细信息

目的:采用人工神经网络模型及在线软件BIMAS和SYFPEITHI共同预测汉滩病毒核蛋白HLA-A)02限制的CTL表位,并对CTL表位进行实验鉴定。方法:①选取2004-11解放军第四军医大学唐都医院住院的肾综合征出血热患者1例,汉滩病毒特异性IgM抗体为阳性。无菌抽取肾综合征出血热患者外周血,分离外周血单个核细胞,液氮冻存备用。②采用JavaNNS软件模拟神经系统对信息的处理过程,建立人工神经网络模型。训练人工神经网络的9肽包括584种HLA-A)02结合肽和130种HLA-A)02非结合肽,随机数字表法分为3部分:80%为训练肽(用来训练模型,使其学习其中隐含的规律),10%为确认肽(防止训练过度,使训练在适当的时候终止),10%为试验肽(对训练好的模型进行初步的评价)。无、低、中、高亲和力9肽训练人工神经网络的输出值分别为0.1,0.4,0.6,0.8。因此,人工神经网络模型的默认阈值为0.4。此外,还应用在线软件BIMAS和SYFPEITHI,共同预测汉滩病毒核蛋白的CTL表位。③采用ELISPOT鉴定汉滩病毒核蛋白特异的HLA-A)02限制的CTL表位。结果:①人工神经网络、BIMAS和SYFPEITHI预测的准确度:3种方法预测的灵敏度分别达到0.89,0.78,0.97,特异性分别为0.90,0.87,0.20。三者共同预测到23个汉滩病毒核蛋白特异的CTL表位。②HLA-A)02限制的CTL表位的实验鉴定:在可刺激肾综合征出血热患者外周血单个核细胞产生阳性反应的3条15肽中,预测到了4个HLA-A)02限制的CTL表位。经ELISPOT实验证实,其中的2种9肽可刺激HLA-A)02阳性的肾综合征出血热患者外周血单个核细胞分泌γ干扰素。结论:肾综合征出血热患者中存在着汉滩病毒特异的CTL应答。汉滩病毒核蛋白CTL表位的有效预测,大幅度减少了合成候选9肽的数量,降低了成本。为肾综合征出血热细胞免疫应答的研究以及多肽疫苗的设计,奠定了重要基础。

关键词：肾综合征出血热表位/分析 HLA—A抗原

来源：评论

学校读者我要写书评

暂无评论

Use of a peptide mimotope to guide the humanization of MRK-16, an anti-P-glycoprotein monoclonal antibody

引用

JOURNAL OF BIOLOGICAL CHEMISTRY 1999年第39期274卷 27371-27378页

作者： Tang, Y Beuerlein, G Pecht, G Chilton, T Huse, WD Watkins, JD Ixsys Inc San Diego CA 92121 USA

A mimotope-guided strategy for engineering antibodies directed against orphan targets or antigens that are difficult to purify was developed and used to humanize the murine MRK-16 monoclonal antibody (mAb), MRK-16 recognizes a conformational epitope of a 170-kDa membrane protein, termed P-glycoprotein (P-gp). Elevated expression of P-gp on tumor cells is associated with resistance to cytotoxic drugs, a major obstacle in chemotherapy. Murine MRK-16 was used to enrich and screen a phage-displayed peptide library to identify reactive mimotopes, One peptide, termed ALR1, was enriched to a greater extent than others and subsequently was expressed as a fusion protein with glutathione S-transferase. ALR1 fusion protein bound MRK-16 specifically and inhibited binding of MRK-16 to cells expressing elevated levels of P-gp. To humanize MRK-16, the murine complementarity determining regions were grafted onto homologous human heavy and light chain variable region frameworks. Framework residues that differed between the murine MRK-16 and the homologous human templates were analyzed and subsequently, five framework positions potentially important for maintaining the specificity and affinity of MRK-16 were identified. A combinatorial library consisting of 32 variants encoding all possible combinations of murine and human residues at the five differing framework positions was expressed in a phage system. In the absence of purified P-gp, ALR1 fusion protein was used as surrogate antigen to screen the antibody library to identify the framework combination that most preserved the binding activity of the mAb. On the basis of the initial screening against the mimotope four antibody variants were selected for further characterization. The binding affinity of these variants for the ALR1 fusion protein correlated with their binding to cells expressing elevated levels of P-gp. Thus, peptide mimotopes which can be identified for virtually any antibody including those that recognize conformati

关键词：氨基酸序列抗体单克隆/化学抗体单克隆/免疫学结合部位抗体结直肠肿瘤表位/分析免疫球蛋白重链/化学免疫球蛋白轻链/化学免疫球蛋白可变区/化学分子序列数据肽库重组融合蛋白质类/免疫学序列比对肿瘤细胞培养的动物人类小鼠

来源：评论

学校读者我要写书评

暂无评论

Development and application of pathovar-specific monoclonal antibodies that recognize the lipopolysaccharide O antigen and the type IV fimbriae of Xanthomonas hyacinthi

引用

APPLIED AND ENVIRONMENTAL MICROBIOLOGY 1999年第9期65卷 4171-4180页

作者： van Doorn, J Ojanen-Reuhs, T Hollinger, TC Reuhs, BL Schots, A Boonekamp, PM Oudega, B Bulb Res Ctr Dept Plant Qual NL-2160 AB Lisse Netherlands Univ Helsinki Dept Biosci Div Gen Microbiol SF-00014 Helsinki Finland Univ Georgia Complex Carbohydrate Res Ctr Athens GA 30602 USA Landbouw Univ Wageningen Dept Nematol NL-6700 ES Wageningen Netherlands Vrije Univ Amsterdam IMBW BCA Fac Biol NL-1081 HV Amsterdam Netherlands

The objective of this study was to develop a specific immunological diagnostic assay for yellow disease in hyacinths, using monoclonal antibodies (MAbs). Mice were immunized with a crude cell wall preparation (shear fraction) from Xanthomonas hyacinthi and with purified type IV fimbriae. Hybridomas were screened for a positive reaction with X. hyacinthi cells or fimbriae and for a negative reaction with X. translucens pv. graminis or Erwinia carotovora subsp. carotovora. Nine MAbs recognized fimbrial epitopes, as shown by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and immunoelectron microscopy;however, three of these MAbs had weak cross-reactions with two X. translucens pathovars in immunoblotting experiments. Seven MAbs reacted with lipopolysaccharides and yielded a low-mobility ladder pattern on immunoblots. Subsequent analysis of MAb 2E5 showed that it specifically recognized an epitope on the O antigen, which was found to consist of rhamnose and fucose in a 2:1 molar ratio. The cross-reaction of MAb 2E5 with all X. hyacinthi strains tested showed that this O antigen is highly conserved within this species. MAb 1B10 also reacted with lipopolysaccharides. MAbs 2E5 and 1B10 were further tested in ELISA and immunoblotting experiments with cells and extracts from other pathogens. No cross-reaction was found with 27 other Xanthomonas pathovars tested or with 14 other bacterial species from other genera, such as Erwinia and Pseudomonas, indicating the high specificity of these antibodies. MAbs 2E5 and 1B10 were shown to be useful in ELISA for the detection of X. hyacinthi in infected hyacinths.

关键词：抗体细菌/免疫学抗体单克隆/免疫学抗体特异性抗原细菌/分析抗原细菌/免疫学酶联免疫吸附测定表位/分析表位/免疫学纤毛细菌/化学纤毛细菌/免疫学免疫印迹法磁共振波谱学显微镜检查免疫电子 O抗原/分析 O抗原/化学 O抗原/免疫学植物疾病/微生物学植物叶/微生物学黄单胞菌属/免疫学动物小鼠

来源：评论

学校读者我要写书评

暂无评论

Organelle isolation by magnetic immunoabsorption

引用

BIOTECHNIQUES 1999年第2期26卷 336-343页

作者： Kausch, AP Owen, TP Narayanswami, S Bruce, BD Univ Tennessee Dept Cellular Biochem & Mol Biol Knoxville TN 37996 USA Connecticut Coll New London CT 06320 USA Jackson Lab Bar Harbor ME 04609 USA Univ Tennessee Knoxville TN USA

Currently most organelle isolation procedures rely on physical parameters and centrifugation for separation. Here. we report the rapid and gentle isolation of a variety of organelles by immunolabeling whole cell lysates with organelle-specific antibodies and streptavidin magnetic particles follow ed by separation in a magnetic field Using magnetic immunoabsorption, we have been able to specifically label mouse metaphase chromosomes mm a variety of plant organelles, including: amyloplasts chloroplasts and nuclei from whole cell lysates of various plant tissues. We find that the distinct magnetic properties, surface characteristics and mean diameter-size ranges of different particle preparations significantly influence their specific utility! for organelle isolations. By using an internal-field magnetic separation device,,we have developed a method for quantitative recovery of labeled organelles in microarrays and rested a variety of antibodies to chloroplast outer envelope proteins for their ability to immune-isolate chloroplasts.

关键词：抗体单克隆生物素细胞分级分离/方法细胞核/化学细胞核/免疫学中心体/化学中心体/免疫学叶绿体/化学叶绿体/免疫学染色体 DNA 植物/分析表位/分析流式细胞术/方法免疫磁化分离/方法原位杂交膜蛋白质类/分析膜蛋白质类/免疫学植物蛋白质类/分析植物蛋白质类/免疫学植物/超微结构抗生蛋白链菌素动物小鼠

来源：评论

学校读者我要写书评

暂无评论

EPITOPE MAPPING BY A METHOD THAT REQUIRES NO AMINO-ACID-SEQUENCE INFORMATION

引用

ANALYTICAL BIOCHEMISTRY 1992年第1期205卷 179-182页

作者： YUAN, J LOW, PS Department of Chemistry Purdue University West Lafayette Indiana 47907 USA

A simple, rapid method of epitope mapping has been developed that avoids the often cumbersome requirement of obtaining amino acid sequence information. The protein antigen is digested with various concentrations of carboxypeptidase into a nearly continuous series of polypeptides of different molecular weights, all containing a common N-terminus. The peptides are separated by polyacrylamide gel electrophoresis and then transferred to nitrocellulose paper. After developing the blot with the antibody to be mapped, a nearly continuous stain is observed extending from the position of the intact antigen to the molecular weight of the smallest N-terminal fragment still containing the antibody's epitope. By noting the molecular weight where the stain terminates, the position of the epitope relative to the N-terminus can be determined. Using this methodology, and taking special precautions to inhibit all endoproteinases in the carboxypeptidase preparation, the previously mapped epitopes of six nonoverlapping antibodies to the erythrocyte anion transporter were confirmed.

关键词：氨基酸序列印迹法蛋白质羧肽酶B 羧肽酶类/免疫学表位/分析分子量胰腺/酶学酵母菌酿酒/酶学酿酒酵母蛋白质类猪动物

来源：评论

学校读者我要写书评

暂无评论

Physical interactions between phospholamban and sarco(endo)plasmic reticulum Ca²⁺-ATPases are dissociated by elevated Ca²⁺, but not by phospholamban phosphorylation, vanadate, or thapsigargin, and are enhanced by ATP

引用

JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第20期275卷 15034-15038页

作者： Asahi, M McKenna, E Kurzydlowski, K Tada, M MacLennan, DH Univ Toronto Banting & Best Dept Med Res Charles H Best Inst Toronto ON M5G 1L6 Canada Merck Res Labs Dept Pharmacol West Point PA 19486 USA Osaka Univ Sch Med Dept Med & Pathophysiol Suita Osaka 5650891 Japan

Previous co-immunoprecipitation studies (Asahi, M,, Kimura, Y., Kurzydlowski, K., Tada, M., and MacLennan, D. H. (1999) J, Biol, Chem. 274, 32855-32862) revealed that physical interactions between phospholamban (PLN) and the fast-twitch skeletal muscle sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA1a) were retained, even with PLN monoclonal antibody 1D11 bound to an epitope lying between PLN residues 7 and 17, Because the 1D11 antibody relieves inhibitory interaction between the two proteins, it was of interest to determine whether PLN phosphorylation or elevation of Ca2+, which also relieves inhibitory interactions between PLN and SERCA, would disrupt physical interactions. Co-immunoprecipitation was measured in the presence of increasing concentrations of Ca2+ or after phosphorylation of PLN by protein kinase k Physical interactions were dissociated by elevated Ca2+ but not by PLN phosphorylation, The addition of ATP enhanced interactions between PLN and SERCA. The further addition of vanadate and thapsigargin, both of which stabilize the E-2 conformation, did not diminish binding of PLN to SERCA. These data suggest that physical interactions between PLN and SERCA are stable when SERCA is in the Ca2+-free E-2 conformation but not when it is in the E-1 conformation and that phosphorylation of PLN does not dissociate physical interactions between PLN and SERCA.

关键词：腺苷三磷酸/代谢氨基酸取代抗体单克隆钙/药理学钙结合蛋白质类/化学钙结合蛋白质类/代谢钙转运ATP酶类/化学钙转运ATP酶类/代谢细胞系表位/分析动力学微粒体/酶学诱变定点磷酰化蛋白质构象重组蛋白质类/化学重组蛋白质类/代谢肌浆网/酶学毒胡萝卜内酯/药理学转染钒酸盐类/药理学人类

来源：评论

学校读者我要写书评

暂无评论

Spatially addressed synthesis of amino- and amino-oxy-substituted 1,3,5-triazine arrays on polymeric membranes

引用

JOURNAL OF COMBINATORIAL CHEMISTRY 2000年第4期2卷 361-369页

作者： Scharn, D Wenschuh, H Reineke, U Schneider-Mergener, J Germeroth, L Humboldt Univ Klinikum Charite Inst Med Immunol D-10098 Berlin Germany Jerini Bio Tools GMBH D-12489 Berlin Germany ChemotopiX GMBH D-12489 Berlin Germany

Effective spatially addressed parallel assembly of trisamino- and amino-oxy-1,3,5-triazines was achieved by applying the SPOT-synthesis technique on cellulose and :polypropylene membranes. In addition to developing a suitable linker strategy and employing amines and phenolate ions as building blocks, a highly effective microwave-assisted nucleophilic substitution procedure at membrane-bound monochlorotriazines was developed. The 1,3,5-triazines obtained could be cleaved in parallel from the solid support by TFA Vapor to give compounds adsorbed on the membrane surface in a conserved spatially addressed format for analysis and screening. The reaction conditions developed were employed for the synthesis of 8000 cellulose-bound 1,3,5-triazines which were probed in parallel for binding tb the anti-transforming growth factor-ct monoclonal antibody Tab2 in order to identify epitope mimics.

关键词：胺类氨基酸序列抗体单克隆药物设计酶联免疫吸附测定/方法表位/分析膜人工分子序列数据氧化物肽类/化学聚合物构效关系转化生长因子α/化学转化生长因子α/免疫学三嗪类/化学合成三嗪类/化学

来源：评论

学校读者我要写书评

暂无评论

CONSTRAINTS ON MODELS FOR THE FOLDING OF THE NA,K-ATPASE

引用

ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 1992年第1期671卷 217-227页

作者： SWEADNER, KJ ARYSTARKHOVA, E HARVARD UNIV SCH MEDDEPT CELLULAR & MOLEC PHYSIOLBOSTONMA 02115

We have attempted to bring together in graphic fashion the available evidence on the structure of the Na,K-ATPase and the H,K-ATPase. There appears to be much room for modification of the existing models for transmembrane folding. More sites on each side of the membrane need to be identified. Whether these will be antibody epitopes, sites of covalent modification, or tags inserted by mutagenesis is less important than that there be many of them and that each be verified by alternative approaches. If any single principle has emerged from the study of the topography of membrane proteins, it is that it is easy to reach conclusions too soon.

关键词：氨基酸序列结合部位抗体钙转运ATP酶类/化学细胞膜/酶学细胞膜/超微结构表位/分析 H(+)K(+)交换ATP酶/化学模型结构分子序列数据蛋白质折叠序列同源性氨基酸钠钾交换ATP酶/化学动物大鼠

来源：评论

学校读者我要写书评

暂无评论

DISTRIBUTION OF ANTIGENIC DETERMINANTS BETWEEN ACTINOMYCES-VISCOSUS AND ACTINOMYCES-NAESLUNDII

引用

JOURNAL OF DENTAL RESEARCH 1988年第1期67卷 15-20页

作者： FIRTEL, M FILLERY, ED UNIV TORONTO FAC DENT 124 EDWARD ST TORONTO M5G 1G6 ONTARIO CANADA

A total of 18 monoclonal antibodies was raised against whole cells of Actinomyces viscosus and Actinomyces naeslundii. The monoclonal antibodies were used to determine the cross-reacting patterns among 26 strains of these species. Eleven different antigenic determinants were found. The specificity profiles of the antibodies indicated that the antigenic determinants of A. viscosus and A. naeslundii were arranged in a complicated mosaic. Extensive cross-reactions occurred between A. viscosus strains and strains of "typical" and "atypical" A. naeslundii. However, cross-reactions were rare between the two groups of A. naeslundii. A. viscosus appears to occupy a "middle position" between the two A. naeslundii groups. In addition to their value in seroclassification, some of the monoclonal antibodies were found to be useful in the identification of these species. One monoclonal antibody appeared to be selective for the "typical" A. naeslundii group. A. viscosus and "atypical" A. naeslundii-specific antibodies were also found, though they did not label every strain in their respective clusters. A. viscosus detection might be improved if mixtures of monoclonal antibodies were used.

关键词：放线菌属/分类放线菌属/免疫学放线菌黏性/分类放线菌黏性/免疫学抗体单克隆抗体特异性交叉反应表位/分析荧光抗体技术间接血清分型/方法种特异性人类

来源：评论

学校读者我要写书评

暂无评论

Elevated soluble MUC1 levels and decreased anti-MUC1 antibody levels in patients with multiple myeloma

引用

BLOOD 2000年第9期96卷 3147-3153页

作者： Treon, SP Maimonis, P Bua, D Young, G Raje, N Mollick, J Chauhan, D Tai, YT Hideshima, T Shima, Y Hilgers, J von Mensdorff-Pouilly, S Belch, AR Pilarski, LM Anderson, KC Dana Farber Canc Inst Div Hematol Malignancies Dept Adult Oncol Boston MA 02115 USA Harvard Univ Sch Med Dept Med Boston MA 02115 USA Bayer Diagnost Walpole MA USA Vrije Univ Amsterdam Acad Hosp Amsterdam Netherlands Cross Canc Inst Dept Med Oncol Edmonton AB T6G 1Z2 Canada

Soluble MUC1 (sMUC1) levels are elevated in many MUC1(+) cancers. We and others have shown that MUC1 is expressed on multiple myeloma (MM) plasma cells and B cells. In this study, we measured sMUC1 levels in bone marrow (BM) plasma from 71 MM patients and 21 healthy donors (HDs), and in peripheral blood (PB) plasma from 42 MM patients and 13 HDs using an immunoassay that detects the CA27.29 epitope of MUC1. sMUC1 levels were found to be significantly greater (mean 31.76 U/mL, range 5.69 to 142.48 U/mL) in MM patient BM plasma versus HD BM plasma (mean 9.68 U/mL, range 0.65 to 39.83 U/mL) (P < .001). Importantly, BM plasma sMUC1 levels were related to tumor burden because sMUC1 levels were significantly higher for NIM patients with active disease (34.62 U/mL, range 5.69 to 142.48 U/mL) versus MM patients with minimal residual disease (16.16 U/mL, range 5.7 to 56.68 U/mL) (P = .0026), sMUC1 levels were also elevated in the PB plasma of MM patients (32.79 U/mL, range 4.15 to 148.84 U/ml) versus HDs (18.47 U/ml, range 8.84 to 42.49) (P = .0052). Lastly, circulating immunglobulin M (IgM) and IgG antibodies to MUC1 were measured in 114 MM patients and 31 HDs, because natural antibodies to MUC1 have been detected in patients with other MUC1-bearing malignancies. These studies demonstrated lower levels of circulating IgM (P < .001) and IgG (P = .078) antibodies to MUC1 in MM patients compared with HDs. Our data therefore show that in MM patients, sMUC1 levels are elevated and correlate with disease burden, whereas anti-MUC1 antibody levels are decreased. (C) 2000 by The American Society of Hematology.

关键词：自身抗体/分析自身抗体/血液活组织检查针吸骨髓/免疫学骨髓/病理学 CA-15-3抗原/分析 CA-15-3抗原/血液细胞培养的表位/分析霍奇金病/血液霍奇金病/免疫学霍奇金病/病理学多发性骨髓瘤/血液多发性骨髓瘤/免疫学多发性骨髓瘤/病理学肿瘤残余/血液肿瘤残余/免疫学肿瘤残余/病理学复发参考值肿瘤细胞培养的肿瘤标记生物学/分析肿瘤标记生物学/血液人类

来源：评论

学校读者我要写书评

暂无评论

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

请选择保存的检索档案：

请选择收藏分类：

通借通还

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：