FITC标记的抗Brdu单克隆抗体的研制

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…     

rhIL-1ra减轻新生大鼠窒息后肾损伤的作用机制

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－     

R-藻红蛋白荧光标记单克隆抗体的研制

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     

FITC-抗FITC系统在夹心法ELISA检测IFN-γ和IL-4中的应用

异硫氰酸荧光黄(FTTC)是一种具有抗原性的荧光素分子，极易与免疫球蛋白形成稳定的结合物，并且不影响抗体结合抗原的活性。偶联了辣根过氧化物酶(HRP)的高效价的抗FITC分子单克隆抗体(mAb)，与FITC标记的特异性一抗组成的FITC-抗FTTC放大系统，可以有效地提高酶联免疫吸附试验(ELISA)的敏感性、特异性，     

两种用FITC标记红桂木凝集素方法的比较

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。     

放射性核素标记的单克隆抗体（McAb）定量测定的一种方法

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   

用单克隆抗体检测肿瘤标记CA19-9的临床研究

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。     

制备单克隆抗体检测PES1的表达

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。     

^99mTc标记单克隆抗体Au14—1对荷宫颈癌小鼠的放射免疫显像 …

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     

抗淋巴样增强因子-1羧基端单克隆抗体的制备及其表位的初步鉴定

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.     

单克隆抗体在肿瘤治疗中的应用进展

近年来用于肿瘤治疗的单克隆抗体药物发展迅速.单克隆抗体与肿瘤细胞上特异性的抗原分子结合,通过抗体依赖性细胞介导的细胞毒作用和补体依赖性细胞毒作用等机制实现杀灭肿瘤细胞的目的.临床抗肿瘤单抗的靶分子主要包括表皮生长因子受体、血管内皮生长因子和部分白细胞分化抗原,用于淋巴瘤、自血病、乳腺癌、直结肠癌、肺癌等领域的治疗已取得瞩目的疗效.     

A monoclonal antibody detecting dipeptidylpeptidase IV in human tissue

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     

A monoclonal antibody against glycoproteins of Aspergillus fumigatus shows anti-adhesive potential

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.     

A novel monoclonal antibody effective against lethal challenge with swine-lineage and 2009 pandemic H1N1 influenza viruses in mice

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.     

Expression of multiple forms of IL-1 receptor antagonist (IL-1ra) by human retinal pigment epithelial cells: identification of a new IL-1ra exon

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.     

LFA-1/ICAM-1单抗对ConA诱导脾淋巴细胞增殖及分泌细胞因子的影响

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。     

131I－抗VEGF单克隆抗体放射免疫治疗荷人骨肉瘤裸鼠的实验研究

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     

单克隆抗体2A10在蛋白芯片技术检测黄病毒属病毒中的应用

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.     

新生血管特异抗体的制备及生物活性鉴定

目的 研制识别肿瘤新生血管的单克隆抗体及其活性。方法 用肝癌细胞条件培养基刺激人血管内皮细胞，以此作为抗原免疫小鼠制备抗体。用免疫组化、ELISA和流式细胞术筛选肿瘤血管特异抗体并鉴定其免疫特性。用MTT法研究抗体对血管内皮细胞增殖的影响。结果 抗体AA98选择性地识别肿瘤血管和人血管内皮细胞，抑制血管内皮细胞的增殖。结论 抗体AA98在肿瘤诊断和治疗方面具有潜在的应用价值。