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1.
FITC标记的抗Brdu单克隆抗体的研制 总被引:3,自引:1,他引:2
利用溴化脱氧尿嘧啶(Brdu)作为胸苷类似物可掺入到新合成的DNA中的特点,通过检测Brdu标记的细胞便能定性、定量地反映细胞的增殖状态〔1〕。我们已筛选出分泌特异性抗Brdu单克隆抗体(mAb)的杂交瘤细胞株〔2〕。为制备荧光素标记的抗BrdumAb用于流式细胞术定量检测增殖细胞,参考有关文献,用亲和层析纯化小鼠IgG亚类的方法,从腹水中纯化了抗BrdumAb,并用FITC荧光素进行标记。经初步用于体外检测Brdu标记细胞的数量,取得较好的结果。1材料和方法1.1材料BXSB小鼠由本校第三临床医学院… 相似文献
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rhIL-1ra减轻新生大鼠窒息后肾损伤的作用机制 总被引:2,自引:1,他引:1
目的 探讨重组的人白细胞介素1-受体拮抗剂(rhIL-1ra)减轻新生大鼠窒息后肾损伤的作用及机制。方法 Witsar大鼠30只,7~10d龄,随机均分为窒息组、窒息+rhIL-1ra处理组和对照组,在窒息30min复氧120min时,测定左肾系数(LRC);用硫代巴比妥酸法测定肾组织过氧化脂质(LPO)含量,用酶联免疫吸附法(ELISA)测定肿瘤坏死因子-α(TNF-α)和白细胞介素-8(IL- 相似文献
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R-藻红蛋白荧光标记单克隆抗体的研制 总被引:4,自引:1,他引:4
目的 研究国产R-藻红蛋白标记二抗的工艺,对异双功能试剂的用量进行比较实验。方法 不同稀释度的异双功能试剂SMCC处理R-PE使之衍生化,SPDP与二抗反应使之衍生化再经DTT处理从而给二抗引入外源巯基。将二者混合反应完成交联。结果 SMCC与R-PE的摩尔比为56:1时,交联物的R-PE与抗体结合比接近1:1,R-PE与抗体的利用率最高。结论 以本文所述工艺进行藻红蛋白的荧光标记,交联物荧光强度 相似文献
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目的:红桂木凝集素(Artocarpus lingnanensis lectin,ALL)与异硫氰酸荧光素(FITC)偶联方法的探究。方法:分别采用透析法和固化法使FITC与红桂木凝集素偶联,检测偶联物的F/P值。结果:固相化法的F/P值达1.02,其它方法 F/P值约为0.758。结论:采用固化法使FITC标记红桂木凝集素,实验重复性较好,方法稳定可靠,且可获得较高偶联率的红桂木凝集素偶联异硫氰酸荧光素(ALL-FITC)利于后续实验研究。 相似文献
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本文用标记McAb与靶细胞结合实验的放射性分析法,为标记单抗的定量提供一个方法,现介绍如下。 标记McAb定量的实验设计 在标记McAb与靶细胞结合反应实验中,每个实验管内液体体积相同,所用的靶细胞恒定在过量的适当浓度(C),标记McAb浓度(T)变化时,根据质量作用定律,McAb与抗原反应处于平衡时得公式Ka=B/(C—B)F(1),Ka为亲和常数,B为与抗原结合的标记抗体,F为游离的标记抗体浓度。整理(1)得KaCF—KaBF=B(2)。(2)两边除以KaCB整理后得F/B=1/KaC+F/C(3)。以F=T—B代入式(3)整理得T/B=1+1/KaC+T/C-B/C(4)。因T相似文献
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用单克隆抗体检测肿瘤标记CA19-9的临床研究 总被引:1,自引:0,他引:1
用单克隆抗体(MeAb)技术在医学各个领域的广泛应用,使肿瘤的免疫学诊断和病情监测出现新的飞跃,同时也发现一些新的肿瘤标记。本文综述用McAb检测近年来发现的肿瘤标记CA19-9的临床研究。 相似文献
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制备单克隆抗体检测PES1的表达 总被引:1,自引:0,他引:1
目的:制备PES1单克隆抗体(mAb)并用所制备的抗体检测PES1在多种肿瘤细胞中的表达及其在成年大鼠不同组织中的表达分布。方法:纯化GST-PES1(1-322aa)融合蛋白后注入小鼠进行免疫,经过细胞融合、筛选以及用Western blot进行特异性鉴定后获得mAb。再用制备的mAb检测PES1在不同的人肿瘤细胞和大鼠不同组织中的表达。结果:所制备的PES1 mAb能特异地识别PES1。用所制备的mAb检测发现,在所检测的人乳腺癌、卵巢癌、肝癌以及肺癌等细胞中均有不同程度的表达;PES1类似物pescadillo在大鼠的乳腺和卵巢组织中表达,但在其他组织中没有被检测到。结论:成功地制备了PES1的mAb。PES1可能与肿瘤的发生发展有一定的关系;由于PES1明显表达于雌激素重要的靶器官,所以可能参与雌激素信号通路。 相似文献
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^99mTc标记单克隆抗体Au14—1对荷宫颈癌小鼠的放射免疫显像 … 总被引:1,自引:0,他引:1
目的和方法:应用^99mTc直接法标记抗小鼠宫颈癌(U14)单克隆抗体Au14-1,对荷Km小鼠进行放射性分布和放射免疫显像的实验观察,探讨用^99mTc-Au14-1作宫颈癌导向诊断和导向治疗的可能性。结果:(1)^99mTc-Au14-1注射后12h肿瘤灶已有放射性聚集,24h肿瘤组织的%ID/g值达8.79,呈明显特异性浓聚;(2)除肾脏外,各脏器的T/NT值在2.02~6.71之间,SPE 相似文献
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目的 制备抗淋巴样增强因子-1(LEV-1)羧基端86个氨基酸(Lct86)特异性单克隆抗体,并初步明确其表位,为进一步研究LEF-1 C端特异序列的功能奠定基础.方法 从Jurkat细胞株中扩增LEF-1 C端特异序列eDNA,并克隆入PQE40、pET32a原核表达载体;分别转化入M15、BL21(DE3)感受态菌,IPIG诱导表达;纯化PQFAO-Lct86融合蛋白作为免疫原免疫Balb/c小鼠制备单克隆抗体,以pET32a-Lct86融合蛋白鉴定单克隆抗体特异性.LEF-1 C端特异序列不同长度的基因片段定向克隆人PQE40,鉴定各片段的表达,利用Western blotting初步鉴定单克隆抗体的表位.结果 成功构建了PQE40-Lct86、pET32a-Lct86重组原核表达载体;纯化获得PQE40-Lct86融合蛋白;得到一株特异性的抗LEF-1 C端单克隆抗体TMAb1,证实其表位位于437aa~454aa.结论 成功制备了一株特异性的抗LEF-1 C端单克隆抗体TMAb1,并鉴定其抗原识别表位位于437aa~454aa,为进一步研究LEF-1 C端的功能奠定基础. 相似文献
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Alfred C. Feller Heinz-Joachim Radzun Eberhard Heymann Helmut Haas Wolfgang Scholz Mohammad R. Parwaresch 《Virchows Archiv : an international journal of pathology》1986,409(2):263-273
Summary Dipeptidylpeptidase IV (DPP IV) occurs among others in exocrine epithelia, hepatocytes, renal tubuli, endothelia, and myofibroblasts of man and laboratory animals. Also Tµ lymphocytes and their varying differentiated neoplastic counterparts reveal this enzyme activity. The present paper describes a new monoclonal antibody recognizing DPP IV.Additional efforts have been taken to detect the subcellular localization of DPP IV and its isoelectric focusing pattern in different tissue types. The monoclonal antibody anti-DPP IV (clone II-19) shows a reaction pattern indistinguishable from the corresponding enzymehistochemical reaction. These findings were further substantiated by immunoblotting analysis. In line with the results of direct enzyme measurements in different subcellular fractions a considerable portion of the enzyme is localized in the membrane fraction.Dedicated to Professor Dr.Dr. h.c. Karl Lennert, Kiel, on the occasion of his 65th birthdayThis study was supported by the Deutsche Forschungsgemeinschaft, SFB 111, program CL1 相似文献
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There has been an increase in the cases of fungal resistance against many antifungal drugs and an effective alternative mode in the form of immunotherapy is being considered as new hope. The adhesion of Aspergillus fumigatus conidia to the host components is one of the prime factors to cause aspergillosis. Carbohydrate components or glycoproteins present on the cell surface play an important role in interaction of the organism to the host and leads to adhesion. Any substance which is capable of disrupting this interaction may be a vital tool for the fungal clearance and hence may protect the host from infections caused by the fungus. In this study, a murine monoclonal antibody IgM generated against the secretory antigens of A. fumigatus, was found to be specific to a common epitope containing glyco-moieties of the various proteins and exhibited anti-adhesive potential in vitro. 相似文献
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The HA protein of the 2009 pandemic H1N1 viruses (H1N1pdm) is antigenically closely related to the HA of classical North American swine H1N1 influenza viruses (cH1N1). Since 1998, through mutation and reassortment of HA genes from human H3N2 and H1N1 influenza viruses, swine influenza strains are undergoing substantial antigenic drift and shift. In this report we describe the development of a novel monoclonal antibody (S-OIV-3B2) that shows high hemagglutination inhibition (HI) and neutralization titers not only against H1N1pdm, but also against representatives of the α, β, and γ clusters of swine-lineage H1 influenza viruses. Mice that received a single intranasal dose of S-OIV-3B2 were protected against lethal challenge with either H1N1pdm or cH1N1 virus. These studies highlight the potential use of S-OIV-3B2 as effective intranasal prophylactic or therapeutic antiviral treatment for swine-lineage H1 influenza virus infections. 相似文献
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Gertjan M. Holtkamp Alex F. de Vos Aize Kijlstra Ron Peek 《European journal of immunology》1999,29(1):215-224
The eye is considered an immunologically privileged organ and is separated from the rest of the body by blood-ocular barriers. Part of the blood-retina barrier consists of the retinal pigment epithelium (RPE). In addition to the physical barrier which the monolayer of RPE cells forms, these cells contribute to ocular immune privilege by producing anti-inflammatory molecules that down-regulate potential damaging immune reactions. In this study the mRNA expression of IL-1 receptor antagonist (IL-1ra) by RPE cells was studied in 15 donor-derived cell lines. Expression of both the intracellular and secreted IL-1ra was detected in unstimulated and IL-1β- or phorbol 12-myristate 13-acetate-exposed RPE. Analysis of IL-1ra protein in RPE cell lysates and cell culture supernatants indicated that these cells produce mainly intracellular IL-1ra. No correlation between IL-1ra expression levels and the IL-1ra gene polymorphism could be detected. In addition to the two known intracellular IL-1ra variants (intracellular IL-1ra type I and type II) evidence is provided for the expression of a hitherto unknown splice variant of the IL-1ra mRNA by RPE cells. Expression was not confined to RPE cells and could also be detected in cultured human fibroblasts and macrophages. This variant, which we have tentatively named intracellular IL-1ra type III, encodes a C-terminally truncated protein of only 27 amino acids. 相似文献
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通过研究LFA-1/ICAM-1单抗对ConA诱导的脾淋巴细胞活化增殖及其分泌细胞因子的影响 ,探 讨了LFA-1/ICAM-1分子在T细胞活化过程中所起的共刺激作用。结果表明,单独应用LFA-1α链单抗(M17/4.4.11.9)或ICAM-1单抗(YN1/1.7.4)均不能引起脾淋巴细胞的增殖,但在加入ConA诱导脾淋巴细胞增殖反应的最初8小时内加入LFA-1单抗可以剂量依赖性地抑制ConA诱导的脾淋巴细胞增殖反应及脾淋巴细胞分泌IL-2、IPN-γ,而加入ICAM-1单抗却无此效应。说明LPA-1在ConA诱导的T细胞活化过程中起着重要的共刺激作用,LPA-1通过与除ICAM-1以外的其它配体分子(如ICAM-3)相互识别,提供T细胞活化所必需的共刺激信号。 相似文献
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目的评价^131Ⅰ抗VEGF单克隆抗体(Sc-7269)瘤内注射放射免疫治疗荷人骨肉瘤裸鼠的疗效。方法 相似文献
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目的 本研究拟探讨1株黄病毒属特异的单克隆抗体2A10作为检测抗体在同时检测5种黄病毒属病毒的蛋白芯片中的应用价值.方法 将病毒的特异性单抗作为捕获抗体进行点样,制备用于检测5种黄病毒属病毒的抗体芯片,然后分别加入拟检测的病毒灭活抗原,捕获病毒颗粒后,加入荧光染料CY3标记的2A10检测抗体进行孵育,通过扫描并分析芯片上不同抗体的荧光强度,确定2A10检测抗体的效价和可检测的病毒滴度,并对2A10的检测效果进行评价.结果 蛋白芯片检测结果表明,2A10可作为检测抗体用于对5种黄病毒属病毒的检测,CY3标记的2A10效价为1∶160,以脾传脑炎(TBE)病毒为例,可检测的病毒滴度为1250 PFU.结论 单克隆抗体2A10作为一株黄病毒属特异的单抗,可以用于蛋白芯片技术检测黄病毒属病毒. 相似文献