McAbGB2识别的乳腺癌血清抗原性质研究

对抗人乳腺癌血清抗原单克隆抗体GB₂（McAbGB₂）识别的抗原（McAbGB₂抗原）性质及含量进行了研究．结果表明,McAbGB₂抗原是由糖及蛋白质组成的复合蛋白质,不耐热;McAbGB₂抗原决定簇不存在于铁蛋白及癌胚抗原;蛋白质印边检测表明McAbGB₂抗原有两条区带,分子量分别为116及45ku,分布于血液及癌组织;用McAbGB₂进行血清学分析（ELISA试验）,结果表明：乳腺癌阳性符合率达98％（50／51）,正常人及乳腺良性肿瘤病人的假阳性率分别为6％（3／50）及6．7％（1／15）．McAbGB₂抗原可能是新的乳腺癌相关抗原．     

抗癌胚抗原单克隆抗体在肿瘤免疫显像中的应用

本文简述了放射性同位素单克隆抗体结合物在人体内的特性、代谢及肿瘤免疫显像的特点,并对抗癌胚抗原单抗在各种肿瘤免疫显像中的应用进行了分类介绍。     

抗胶质瘤细胞单克隆抗体SZ-38及其识别抗原的生化特征

本文以人脑胶质瘤细胞系SHG-44为抗原,利用杂交瘤技术建立了一株恒定地分泌抗胶质瘤细胞单克隆抗体(McAb)的杂交瘤细胞株SZ-38。McAb SZ-38与9/10胶质瘤细胞系发生强结合反应。而与淋巴细胞、ABO型红细胞等所有正常血液细胞及绝大多数被检测肿瘤细胞系无反应。经免疫转移电泳及免疫沉淀法鉴定,该McAb识别抗原为胶质瘤细胞膜上Mw47,000糖蛋白。应用SPA-Sepharose 4B提纯MeAb SZ-38,再把纯化抗体交联于Sepharose 4B,以McAb亲和层析法提取SZ-38抗原,经SDS-PAGE证实其有较高的纯度。     

抗人肺癌细胞单克隆抗体的制备及其特性的研究

用人肺鳞癌细胞LTEP-78细胞系免疫Blab/c小鼠获得3株抗人肺癌细胞的单克隆抗体杂交瘤系。其中BLTI-01株经六次克隆化培养,体外传代8个月以上。BLTI-01与白细胞抗原及血型抗原基本上无交叉反应;与骨髓细胞无交叉反应;与癌胚抗原和胎甲球蛋白不相关;与肺鳞癌、肺腺癌细胞系及部分其它肿瘤细胞呈阳性反应。     

伤寒杆菌特异性抗原的检测与单克隆抗体的应用

<正>一、概述 伤寒病在发展中国家仍然是一个严重的公共卫生问题。近年来,我国局部地区相继发生伤寒暴发流行,强度大、持续时间长、患者病情重、并发症多、伤寒症状不典型,加之出现耐氯霉素的M1型菌株,这就给临床诊断以及预防、治疗和控制疾病带来了新的困难。伤寒病的早期、快速、特异和     

抗胃癌单克隆抗体3H11对应抗原的纯化及鉴定

抗胃癌单克隆抗体3H11的对应抗原主要表达于靶细胞的膜上,不耐热,经蛋白酶消化,抗原失活。过碘酸氧化不影响活性,Schiff′s试剂糖柒色为阴性。SDS-PAGE显示单一区带,分子量为210kD。靶细胞经衣霉素抑制糖基化作用后,对其分子量无明显影响。等电聚焦电泳表明其等电点为8.5,故3H11抗原为一大分子碱性蛋白。     

长叶车前花叶病毒上海分离株单克隆抗体的制备及其免疫学特性 Ⅲ 病毒抗原决定簇的单克隆抗体识别

前文分别报道了长叶车前花叶病毒上海分离侏(RMVsh)单克隆抗体的制备及根据它们在不同免疫反应中的特性,将它们分为两组,分别识别性质不同的抗原决定簇。本文采用修改的Friguet方法测定了各组内各单克隆抗体之间的增值反应(Additivity Reaction)特性,并分析了它们识别抗原决定簇的特性。村料与方法一、病毒及单克隆抗体长叶车前花叶病毒上海分离株及其单克隆抗体1H2、7H1、10H1、11H2、12H3、17H6、29H1来源、制备和特性见前文报道。二、抗原饱和曲线的测定抗原饱和曲线测定采用间接ELISA办法,抗原浓度为2μg/ml。     

Identification of a 220-kDa membrane tumor-associated antigen by human anti-UK114 monoclonal antibodies selected from the immunoglobulin repertoire of a cancer patient

Human monoclonal antibodies (HuMAb) specific for a 14-kDa perchloric acid-soluble protein (defined as UK114) were produced by somatic fusion of the human-mouse myeloma K6H6/B5 with Epstein-Barr virus-transformed peripheral B lymphocytes from a cancer patient previously treated with UK101 preparations, containing the UK114 protein. Three IgM-secreting clones were selected on the criteria of specificity for the purified UK114 protein immobilized onto plastic and adapted to grow in a serum-free medium. The reactivity of these antibodies showed a broad distribution pattern restricted to fresh tumor tissues and tumor cell lines, mainly of the adenocarcinoma type. None of the normal cells, nonmalignant cell lines, and normal tissues surrounding the neoplastic lesions were reactive. The immunochemical analysis of the target antigens showed that the HuMAb recognize a molecule of 220 kDa selectively expressed by the surface of tumor cells, as well as a cytoplasmic 14-kDa protein. The 220-kDa antigen was different from other tumor-associated antigens with similar molecular mass and, so far, unique. In the presence of human complement, two of three HuMAb are cytotoxic for tumor cells expressing the 220-kDa surface antigen. The tumor specificity and the lytic ability attributed to these HuMAb are promising features for the exploration of future clinical applications.     

抗肿瘤单抗3H11对应抗原cDNA片段的克隆

单克隆抗体３Ｈ１１能与多种肿瘤细胞特异结合,克隆其对应抗原无疑具重要意义．用胃癌细胞ＭＧＣ８０３构建ｃＤＮＡ表达文库,通过抗体３Ｈ１１对其进行原核表达筛选,获得一株能与３Ｈ１１特异反应的阳性克隆．其ｃＤＮＡ插入片段为５５４ｂｐ．ＧｅｎＢａｎｋ不含其同源序列．将此ｃＤＮＡ片段与谷胱甘肽转移酶表达质粒ｐＧＥＸ－４Ｔ重组,Ｗｅｓｔｅｒｎｂｌｏｔ和竞争抑制实验表明,表达产物依然保持同３Ｈ１１反应的特异性．可见它是３Ｈ１１对应抗原的ｃＤＮＡ．     

Detection of Human Leukocyte Antigen Biomarkers in Breast Cancer Utilizing Label-free Biosensor Technology

According to the American Cancer Society, more than 200,000 women will be diagnosed with invasive breast cancer each year and approximately 40,000 will die from the disease. The human leukocyte antigen (HLA) class I samples peptides derived from proteasomal degradation of cellular proteins and presents these fragments on the cell surface for interrogation by circulating cytotoxic T lymphocytes (CTL). Generation of T-cell receptor mimic (TCRm) monoclonal antibodies (mAbs) which recognize breast cancer specific peptide/HLA-A*02:01 complexes such as those derived from macrophage migration inhibitory factor (MIF_19-27) and NY-ESO-1_157-165 enable detection and destruction of breast cancer cells in the absence of an effective anti-tumor CTL response. Intact class I HLA/peptide complexes are shed by breast cancer cells and represent potentially relevant cancer biomarkers. In this work, a breakthrough biomarker screening system for cancer diagnostics incorporating T-cell receptor mimic monoclonal antibodies combined with a novel, label-free biosensor utilizing guided-mode resonance (GMR) sensor technology is presented. Detection of shed MIF/HLA-A*02:01 complexes in MDA-MB-231 cell supernatants, spiked human serum, and patient plasma is demonstrated. The impact of this work could revolutionize personalized medicine through development of companion disease diagnostics for targeted immunotherapies.     

Novel tumor-associated antigen of hepatocellular carcinoma defined by monoclonal antibody E4-65

A monoclonal antibody, E4-65, produced by immunizing mice with SMMC-7721 cells, a human hepatocellular carcinoma (HCC) cell line, was used to identify and characterize an unreported HCC-associated antigen. Indirect immunofluorescence studies showed that E4-65 antibody reacted with five out of eight HCC cell lines, but not with 10 non-HCC tumor cell lines or a normal liver cell line. Using immunohistochemical examination, E4-65 antigen was detected on the cell membranes and in the cytoplasm of human liver tumor tissues, but was not found in most other tumors, or normal adult or fetal tissues, except for a weakly positive reaction in tissues of the digestive system. Western blot analysis showed that E4-65 antibody bound to a 45 kDa protein in the human HCC cell line and tissue lysates. Enzyme treatment and lectin blotting did not detect the carbohydrate chain in E4-65 antigen. This HCC-associated protein represents a potentially useful target for diagnoses and immunotherapy of human HCC.     

Mapping and Characterization of a Sequential Epitope on the Rabies Virus Glycoprotein Which Is Recognized by a Neutralizing Monoclonal Antibody,RG719

We have established a murine hybridoma cell line RG719 which produces a rabies virus-neutralizing IgM-type monoclonal antibody (referred to as MAb RG719). Immunoblot analysis indicated that the antibody recognized a sequential epitope of G protein. Among four rabies virus strains tested, the antigenicity to MAb RG719 was absent from the Nishigahara strain, while the other three strains (HEP, ERA and CVS) reacted to the MAb. Studies with deletion mutants of the G protein indicated that the epitope was located in a middle region of the primary structure of G protein, ranging from position 242 to 300. By comparing the estimated amino acid sequence of the four strains, we found in this region two amino acids (at positions 263 and 291) which are common to three of those strains but are not shared by the Nishigahara strain. The site-directed point mutagenesis revealed that replacement of phenylalanine-263 by leucine destroyed the epitope of the HEP G protein, while the epitope was generated on the Nishigahara G protein whose leucine-263 was replaced by phenylalanine. These observations suggest that phenylalanine-263 is essential for constructing the epitope for MAb RG719. The synthetic 20-mer peptide produced by mimicking the amino acid sequence (ranging from amino acid positions 249 to 268) of the presumed epitope region was shown to bind specifically to MAb RG719 and also to raise the virus-neutralizing antibodies in rabbits. Vaccination with the HEP vaccine produced in Japan induced in humans and rabbits production of significant amounts of the antibodies which reacted with the 20-mer peptide.     

CHO细胞表达的抗炭疽保护性抗原人源化抗体的纯化及质量控制分析

目的:建立CHO细胞表达的抗炭疽保护性抗原人源化单抗纯化工艺和质量控制方法。方法:收获50 L生物反应器中无血清悬浮培养的CHO工程细胞培养液,通过高速离心去除细胞及碎片后超滤浓缩上清液,经亲和层析、SPFF阳离子交换层析后,将所得目的蛋白质经G25凝胶柱更换缓冲液以完成纯化;对纯化的产品进行单抗鉴别(Western印迹)、相对分子质量(SDS-PAGE和MOLDI-TOF)、纯度(SEC-HPLC)、生物学活性(毒素中和试验)、产品相关杂质(SEC-HPLC检测聚集体、降解产物)、工艺相关杂质(ELISA分析残余宿主蛋白、残余蛋白A)、安全性(凝胶法检测内毒素、薄膜过滤法考察无菌)分析。结果:样品回收率达61.7%;单抗鉴别实验阳性;MOLDI-TOF测定完整分子的相对分子质量为147 995,与预期相符;单体比例为99.25%,二聚体比例为0.75%;EC50值为0.1516μg/m L。残余宿主蛋白、残余蛋白A、内毒素、无菌检查结果符合药典要求。结论:初步建立了纯化工艺和质量控制方法,为抗炭疽人源化单抗药物的研制奠定了基础。     

Further Studies on Guinea Pig Z-1 Antigen That Is Involved in Phagocytosis of Zymosan by Macrophages: Cell Type Distribution of the Antigen and Cross-Reactivity of Anti-Z-1 with Human CR3

We have previously identified a heterodimer molecule, Z-1, on guinea pig peritoneal macrophages (Møs) by the newly prepared monoclonal antibody, anti-Z-1, and Z-1 has been assumed to be the complement receptor type three (CR3) in this species. To clarify this assumption, the cell type distribution of the antigen in guinea pig and the cross-reactivity of anti-Z-1 with other species were analyzed. It was demonstrated that Z-1 was expressed on peritoneal Møs, pulmonary Møs, peritoneal polymorphonuclear leukocytes (PMN), peripheral neutrophils, and some subpopulations of spleen cells and of bone marrow cells, but not on erythrocytes, circulating lymphocytes, and lymphocytes in both spleen and bone marrow in detectable amounts. Thus the expression of Z-1 seems to be restricted to phagocytes as is CR3 of other species. Furthermore, it was found that anti-Z-1 bound with peripheral neutrophils from human, horse and goat and HL-60 cells differentiated into monocytes. Any cross-reactivity of the antibody was not detected with neutrophils from rabbit, cow, sheep and dog and nondifferentiated HL-60 cells. Human Z-1 was indistinguishable from human CR3, since both were the heterodimer consisting of α chain of 170 kDa (pI = 6.6-7.2) noncovalently associated with β chain of 100 kDa (pI = 5.6-6.7). In addition, human CR3 in detergent-lysate of neutrophils was completely adsorbed with anti-Z-1 F(ab')2-Sepharose. These findings indicate that guinea pig Z-1 shares an antigenic determinant with human CR3. It thus seems to be possible that Z-1 may function as CR3 in guinea pigs.     

Determination of Ki67 Growth Fraction and Oestrogen Receptors In Screen-Detected Breast Cancer Using Cytological Preparations

Oestrogen receptor (ER) status of 77 cases of screen-detected breast cancer has been determined using cytological preparations. In 48% ER status was positive, which was the same proportion as that formed in a control group of age-matched patients with symptomatic breast carcinoma. Since the screen-detected group contained more low grade tumours, the percentage of ER-positive cases would be expected to be higher. the reasons for the discrepancy are discussed. Ki67 score has been determined for 41 cases of screen-detected cancer. Ki67 score showed a positive correlation with histological tumour grade and a negative correlation with ER status. However, there was no correlation with tumour size or lymph node status. the Ki67 scores in the screen-detected cancers were essentially similar to those found in an age-matched symptomatic group, but the very low scores were only found in the screened group.