抗乳腺癌单克隆抗体的制备及其应用

目的:制备抗人乳腺癌单克隆抗体(McAb),用于临床肿瘤的免疫病理诊断与分型研究,为药物导向诊断和治疗奠定基础.方法:以乳腺癌细胞系ZR75免疫Balb/c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/0进行融合,经选择培养、筛选及克隆化,获得恒定分泌抗ZR75单克隆抗体的杂交瘤细胞系BM109.结果:单抗BM109的Ig亚类为IgG1,BM109抗原主要表达在靶细胞膜上,为分子量37KD的蛋白质抗原.在乳腺癌的免疫反应率为86.7%(26/30),与胃癌、食管癌、卵巢癌等6种肿瘤的免疫反应率为9.5%(4/42),而对乳腺纤维瘤、囊性增生症和周围正常乳腺组织未见阳性反应.结论:所获一株抗乳腺癌单克隆抗体,具有较好的特异性,可望进一步用于乳腺癌的诊断、治疗及其发生发展的研究.     

抗p185单克隆抗体的制备及其在乳腺癌诊断中的应用

ｐ18 5是癌基因ｃ ｅｒｂＢ 2编码的跨膜蛋白 ,ｃ ｅｒｂＢ 2在乳腺癌等 10余种常见癌症中均有过量表达 ,与癌细胞的生物学行为密切相关[1] 。我们采用修饰的细胞表面区域表位包埋法制备出三株抗ｐ185单克隆抗体 ,并进行了临床验证。一、材料与方法1 细胞系及主要试剂 :ＳＰ2 / 0、ＮＩＨ/ 3Ｔ3细胞株 ,6～ 8周的ＢＡＬＢ/ｃ雌鼠源于中科院细胞库。Ｔ6 17、ＮＥ91、ＳＫＢＲ 3、ＭＣＦ 7均由美国宾州大学医学院ＭａｒｋＧｒｅｅｎｅ教授实验室惠赠。羊抗鼠ＩｇＧ ＨＲＰ购于卫生部上海生物制品公司 ,商品化抗ｐ185多抗、单抗及Ｉｍｍｕｎｏ…     

人源性抗肝癌单克隆抗体的制备及初步鉴定

目的建立产生人源性抗肝癌单克隆抗体的人-鼠杂交瘤.方法用IL-2和美洲商陆(PWM)体外刺激肝癌病人外周血淋巴细胞,再与小鼠骨髓瘤细胞NS-1融合,待杂交瘤生长进行筛选、克隆化及特性分析.结果建立了3株产生人源性抗肝癌单克隆抗体的人-鼠杂交瘤-AF3,CB7、CH1.三种单抗均识别肝癌细胞表面抗原,并具有显著的补体依赖的细胞毒活性.其中AF3的效价最高.结论通过人-鼠杂交瘤技术获得了产生人源性抗肝癌单克隆抗体的杂交瘤细胞系.抗体效价高,是一种较理想的单克隆抗体.     

抗人肝癌细胞膜单克隆抗体HCMP—1的制备及其生物学特性的初步鉴定

用超选择免疫法，诱导ＢＡＬＢ／ｃ小鼠对正常人肝细胞抗原产生选择性低兔疫反应后，二步腹腔注射人肝癌细胞悬液，２－４周后用人肝癌细胞作强化免疫动物１次，取出脾细胞与ＳＰ２／０骨髓瘤细胞用ＰＥＧ法融合制备杂交瘤。采用细胞抗原ＥＬＩＳＡ法，同时测定培养上清对肝癌细胞及正常肝细胞的免疫反应，筛出一株能稳定分泌抗人肝癌细胞膜抗原的单克隆抗体的杂交瘤细胞株，ＨＣＭＰ－１ＭＡｂ是一种ＩｇＧ２亚型的单抗，它与ＨＢｓ     

抗精胺单克隆抗体的制备及特性

多胺(polyamine)是一个胺类家族的总称。其主要成员有腐胺(putrescine,简称PUT),精脒(spermidine,简称SPD)和精胺(Spermine,简称SPM),它们都是一类直链小分子有机碱,参与DNA、RNA和蛋白质合成的调控,影响其结构与功能。1971年,Russell等首次发现,肿瘤病人或其它组织增生患者的尿液中多胺水平升高,此后有文献相继报导:癌症病人的血清、尿液及体液中各类多胺水平均升高,近年来开展了多胺作为恶性肿瘤诊断指标的研究。     

抗人乳腺癌单克隆抗体的研究现状

１０年来，肿瘤免疫学家用乳癌细胞系、新鲜乳癌组织及人乳汁球膜作免疫原已制备了数十种抗乳癌单抗。本文综述了具代表性的单抗，分析了各种免疫方式的优缺点，并介绍了与乳腺癌相关的一些单抗，提出今后的发展方向。     

抗人肝癌细胞膜单克隆抗体HCMP－1的制备及其生物学特性的初步鉴定

用超选择免疫法，诱导ＢＡＬＢ／ｃ小鼠对正常人肝细胞抗原产生选择性低免疫反应后，二步腹腔注射人肝癌细胞悬液，２～４周后用人肝癌细胞作强化免疫动物１次，取出脾细胞与ＳＰ２／０骨髓瘤细胞用ＰＥＧ法融合制备杂交瘤。采用细胞抗原ＥＬＩＳＡ法，同时测定培养上清对肝癌细胞及正常肝细胞的免疫反应，筛出一株能稳定分泌抗人肝癌细胞膜抗原的单克隆抗体（ＨＣＭＰ－１ＭＡｂ）的杂交瘤细胞株。ＨＣＭＰ－１ＭＡｂ是一种ＩｇＧ２ａ亚型的单抗，它与ＨＢｓＡｇ、ＨＢｃＡｇ、ＨＢｅＡｇ等乙型肝炎病毒相关抗原无交叉免疫反应，与ＡＦＰ－亦无阳性反应。免疫萤光法证实ＨＣＭＰ－１ＭＡｂ对ＱＧＹ－７７０３、ＢＥＬ－７４０２及ＳＭＭＣ－７７２１等人肝癌细胞株呈阳性反应。改良ＡＢＣ免疫组织化学证明ＨＣＭＰ－１ＭＡｂ与肝细胞癌组织呈阳性反应。说明ＨＣＭＰ－１ＭＡｂ对肝癌反应的阳性率较高，特导性较好。超选择免疫法，先联合应用免疫抑制药物使动物对正常肝细胞抗原产生低免疫反应性。再以肝癌细胞免疫时，有利于动物免疫系统对肿瘤相关抗原的识别及特异性抗体的产生。从而提高了杂交瘤制备的成功率。我们认为这一方法在单抗制备方面有较高的应用价值。     

抗人乳腺癌单克隆抗体BG6在乳腺癌诊断和鉴别诊断中的应用

本文应用抗人乳腺癌单克隆抗体BG6对26例临床与病理疑难的乳腺疾病进行免疫组织化学研究。对乳腺的原发或继发癌的鉴别诊断13例中,阳性11例,定为原发瘤;阴性2例则为继发瘤。在难分良、恶性诊断8例中,阳性6例,确诊恶性,其余2例阴性为良性。而在恶性淋巴瘤3例中皆为阴性,1例阳性确诊为未分化癌。这一结果表明,抗人乳腺癌单克隆抗体BG6除有其敏感性外,还具有一定的特异性和选择性。它不仅在乳腺癌的鉴别诊断中有一定的实用价值,而且还可望进一步用于乳腺肿瘤的免疫病理研究。     

抗人肿瘤细胞核单克隆抗体的制备及鉴定

抗人肿瘤细胞核单克隆抗体的制备及鉴定万文徽，韩燕，张宏，魏淑敏，张梅敏，董志伟北京市肿瘤防治研究所（北京１０００３４）以抗肿瘤细胞膜抗原的单克隆抗体（ＭｃＡｂｓ）为载体的导向治疗虽然在动物实验治疗中取得相当进展，但在人实体瘤的临床研究中，其效用却是颇...     

用单克隆抗体SC3A检测人乳腺肿物组织中的相关抗原

本文以抗人结肠癌相关抗原的ＭＡｂＳＣ３Ａ为探针，用免疫组化法检测了其相应抗原在１５３例１４种不同组织类型乳腺肿物组织中的表达与分布。结果在８０例９型的乳腺癌中有７８例（９７．５％）呈阳性结合，其中５７例（７１．３％）的反应范围在５０％以上，表明这种相关抗原在乳腺癌中有较高的表达，这对乳腺癌的有关研究具有实际应用意义，此抗原在７３例５种乳腺良性病变中亦有４５．２％的阳性表达，提示其与乳腺癌之间可存在     

乳腺癌单链抗体制备及其在荷人乳腺癌裸鼠中的放免显像

目的：制备抗乳腺癌单链抗体,用于荷瘤裸鼠的放免显像研究。方法：提取乳腺癌膜抗原,制备抗乳腺癌单克隆抗体M4G3,用Pharmacia噬菌体单链抗体制备系统得到可溶性具有抗乳腺癌活性的ScFv抗体,利用环DTPA法标记核素99^mTc,进行体外培养的乳腺癌细胞系MCF-7和荷瘤小鼠体内定位显像。结果：免疫组化显示单链抗体与乳腺癌膜抗原有较强的亲和力,并在体外培养的乳腺癌细胞及荷瘤小鼠体内显像成功,且在抗体注射第3-5天显像最清晰。结论：提示M4G3的ScFv可在乳腺癌的放射免疫显像和定位诊断研究中发挥作用,值得进一步研究。     

McAb3D3与平阳霉素交联物的制备及其细胞杀伤实验

用ＤｅｘｔｒａｎＴ４０为中介体的方法交联抗人肺腺癌单抗３Ｄ３和平阳霉素（Ａ５），每克分子抗体可携带６８克分子的药物，交联物中抗体活性仍能达到１０－４水平，能较好地与靶细胞Ｌ－３４２结合。体外细胞毒实验，显示交联物和游离Ａ５对靶细胞５０％杀伤浓度分别为０．９μｍｏｌ／Ｌ和４．６μｍｏｌ／Ｌ，杀伤效果比游离Ａ５强５倍多。单抗对靶细胞几乎没有杀伤作用；无关抗体交联物的作用也很弱。交联物和游离Ａ５对非靶细胞ＭＧｃ－８０３的杀伤作用无明显差别。结果提示ＭＣＡｂ３Ｄ３具有良好的导向作用，能携带结合的Ａ５特异地杀伤肺腺癌Ｌ－３４２细胞，可作为临床抗肿瘤药物导向治疗的一种载体。     

A Monoclonal Antibody, KM10 Reactive with Human Gastrointestinal Cancer and Its Application for Immunotherapy

A monoclonal antibody, KM10 (IgG₁) was produced by fusing spleen cells from a human gastric cancer cell (MKN45)-primed BALB/c mouse with the murine myeloma cell line X63-Ag8-653. The antibody reacted strongly with the plasma membrane of human gastrointestinal carcinoma. Sections of the malignant and benign tissues were tested with immunoperoxidase. All of 10 (100%) large intestinal cancers, 26 of 31 (84%) gastric cancers, 5 of 7 (71%) pancreatic cancers and all of 3 (100%) ampullary cancers reacted positively. Moderate or weak reactivity was observed with normal human tissues, hepatoma and carcinomas of mammary, thyroid and adrenal glands. According to a study of the distribution of ¹²⁵I-labeled KM10 in nude mice bearing human gastric cancer, KM10 selectively localized in tumor tissue rather than normal tissue. Whole body autoradiography also supported such a selective distribution. Destruction of antigenic properties by pronase digestion demonstrated its protein nature and by Western blot analysis, it was identified as a protein with an Mr of 180–200 kd. KM10-adriamycin (ADM) conjugate was prepared via an oxidized dextran bridge and this immunoconjugate retained the binding activity against human gastric cancer. MKN45 cells were inoculated subcutaneously into athymic mice and intravenous treatment was begun when the tumor became measurable. A dose-dependent antitnmor activity was observed in vivo with KM10-ADM conjugate, while this conjugate was less toxic than free ADM.     

胃癌单克隆抗体3H11的应用

胃癌单克隆抗体(McAb)3H11具有高阳性反应率、高选择性及高亲和力的特点.以~(131)I标记后注入荷胃癌裸鼠模型中,其瘤/肝比可达8.26±1.26,定位指数达6.08 ±1.51,ID%/g达11.00±2.62.该McAb与~(131)I偶联后可明显增强后者对肿瘤的杀伤效应及减低毒副反应.~(131)I标记的McAb 3H11静脉注入19例拟行手术的胃癌患者中,16例获阳性显像(84.2%).经胃镜引导注入癌旁粘膜下行胃癌放射免疫导向手术,判别胃壁肿瘤浸润的灵敏度、特异性及准确率分别为946%,967%及959%.判别淋巴结转移的上述指标分别为99.2%、97.7%及98.8%.     

Detection of Locally Recurrent Colorectal Cancer with Radiolabeled Monoclonal Antibody H-15

H-15 (HT-29-15) is an IgG1 mouse monoclonal antibody (mAb) to a cell surface antigen (molecular mass, 200,000 daltons) present on virtually all colorectal cancers and also in normal pancreatic ducts and bile ducts, but not in other normal tissues. The biological distribution and imaging characteristics of iodine-131 (^l31I)-labeled mAb H-15 were studied in 5 primary colorectal cancer patients and 9 patients with local recurrence of colorectal cancer. H-15 mAb labeled with 0.5-10 mCi of ¹³¹I was administered 7 to 8 days before surgery at 4 dose levels, ranging from 0.2 to 6 nig. Selective mAb H-15 localization to tumor tissues was demonstrated in 6 of 12 patients with antigen-positive tumors: in two patients, recurrent tumors were negative to H-15 mAb, although the primary tumors were positive. In six patients with positive radioimaging, tumor:normal tissue ratios ranged from 2.05 to 5.35 and tumor:serum ratios from 1,18 to 2.73. The clarity of images seems to correlate well with the latter ratios. Technetium-99 (^99mTc)-albumin blood pool studies in selected cases showed that local recurrence of colorectal cancers was hypovascular, emphasizing the selective localization of mAb H-15 despite poor blood flow distribution in the tumors. The results altogether demonstrated that radioimmnnodetection with ¹³¹I mAb H-15 is valuable for differentiating recurrent colorectal cancer from granuloma formation after surgery.     

Human Monoclonal Antibody Against Colon Cancer

The purpose of the study was to produce human monoclonal antibodies (hMcAb) against human colon cancer for use in radioimmunoimaging. Human-mouse heterohybridomas were developed by fusing SHM-D33 mouse-human hybrid heteromyeloma cells with human lymphocytes from colon cancer tumor-draining lymph nodes. The hybridomas capable of secreting human monoclonal antibodies were screened by using human colon cancer cell lines and pathological biopsies with ELISA and immunohistochemical methods. hMcAb clone H11 was selected for a large-scale antibody production, which was purified from mouse ascites. Biodistribution study demonstrated that specific uptake of ¹²⁵I-hMcAb H11 by human colon cancer xenografts was significantly higher than by normal tissues. Radioimmunoimaging of human colon cancer xenografts exhibited distinct tumor visualization during the period of 72-96 hr after intraperitoneal injection of ¹²⁵I-hMcAb H11. The development of human monoclonal antibodies such as hMcAb H11 may be useful for radioimmunodetection and therapy of colon cancer.     

单克隆抗体MG9用于肺癌的免疫组化研究

用鼠抗人胃癌单克隆抗体MG_9对147例不同组织学类型的肺癌进行了免疫组织化学研究,探讨MG_9相关抗原在肺癌组织中的表达及其临床意义。结果表明,肺癌MG_9相关抗原表达总阳性率为51.7％(76/147);不同组织学类型肺癌间阳性率差异显著,其中腺癌阳性率92.1％(47/51),显著高于鳞癌的30.2％(13/43)和小细胞癌的30.2％(16/53),P<0.001。这提示:单抗MG_9对肺腺癌有较高特异性,可作为肺腺癌的一种新的标志物。