应用抗甲胎蛋白单克隆抗体对AFP抗原性的研究(摘要)

<正> 应用淋巴细胞杂交瘤技术制备出11株抗人AFP单克隆抗体。根据抗不同决定簇的单克隆抗体与其相应抗原结合过程中不相互竞争的原理,应用交叉二位夹心固相放射免疫分析法,鉴别出AFP分子上至少有四个不同的抗原决定簇。一种单克隆抗体与胃蛋白酶分解的一个片断特异性结合。两种单克隆抗体与小牛     

抗华支睾吸虫单克隆抗体杂交瘤细胞株的建立及其初步应用

本文用华支睾吸虫成虫可溶性抗原免疫的BALB/c小鼠与SP_2/o骨髓瘤细胞融合,经筛选和克隆化后获得4株抗华支睾吸虫单克隆抗体杂交瘤细胞。该4株荜克隆抗体与华支睾吸虫成虫抗原作ELISA,结果均为阳性。经交叉试验(ELISA法)证明与日本血吸虫成虫抗原,虫卵抗原和卫氏并殖吸虫成虫抗原均呈阴性反应。4株单克隆抗体免疫球蛋白亚类鉴定除3F_7株为IgG_1外,其余3株均属IgM。初步试用3F_7株单克隆抗体作ELISA双抗体法检测人体华支睾吸虫循环抗原。     

一种制备抗复合抗原单克隆抗体的方法

目的 建立通过单次细胞融合制备抗复合抗原(多种抗原)单克隆抗体的方法。方法 用复合抗原(甲胎蛋白、癌胚抗原、乙型肝炎表面抗原、乙型肝炎核心抗原、乙型肝炎e抗原)免疫BALB/c小鼠,采用常规杂交瘤技术制备单克隆抗体。通过Protein-G亲和层析纯化阳性杂交瘤细胞株腹水,并通过亚类试剂盒法和非竞争ELISA法分别测定抗体亚类和亲和常数,同时应用免疫印迹分析其特异性。结果 通过单次细胞融合共获得20株抗上述复合抗原的单克隆抗体,其中针对甲胎蛋白的单克隆抗体5株、针对癌胚抗原的单克隆抗体6株、针对乙型肝炎表面抗原的单克隆抗体3株;针对乙型肝炎核心抗原的单克隆抗体4株、针对乙型肝炎e抗原的单克隆抗体3株。已鉴定的部分阳性杂交瘤细胞株亚类均为IgG1,抗体亲和常数介于1×10⁹M^-1~2.8×10¹¹M^-1,免疫印迹分析显示,所获得的单抗都与其抗原发生特异性结合。结论 建立了通过单次细胞融合制备针对复合抗原的单克隆抗体制备技术,使单克隆抗体的产出率显著提高,制备周期明显缩短,为建立规模化单克隆抗体制备平台奠定基础。     

胃癌单克隆抗体MGc1相应抗原的初步研究

作者报告一株鼠抗人胃癌单克隆抗体MGcl相应抗原的纯化及分析。MGcl经50％饱和硫酸铵盐析,DEAE-52纤维素柱层析后,获得纯化的IgG 56.31 mg与2 g溴化氰活化的 sepharose 4B偶联,制备免疫吸     

单克隆抗体检测血吸虫卵循环抗原的研究

单克隆抗体检测血吸虫卵循环抗原的研究＊郑南才１蒋作君１徐镜２王维１袁有赉２Ｅ１２株杂交瘤为本室研制，其分泌的单克隆抗体所针对靶抗原为日本血吸虫虫卵分泌性抗原，为了进一步验证此单克隆抗体的应用价值，进行了下列实验。１材料与方法１．１单克隆抗体制备以１０...     

一种新的胃癌相关糖脂糖蛋白抗原的研究

报告一株鼠抗人胃癌单克隆抗体(单抗)MG7相应抗原的纯化及鉴定。单抗MG7经盐析沉淀,DEAE-52纤维素柱层析纯化后,与溴化氰活化的Sepharose 4B偶联,制备免疫吸附剂,用免疫亲和层析的方法从胃癌组织可溶性抗原提取液中纯化出MG7相应抗原。经高效薄层色谱,TLC放射免疫自显影,SDS-聚丙烯酰胺凝胶电泳及免疫印渍等实验分析,证实MG7相应抗原含有中性糖脂和糖蛋白,抗原决定簇存在于糖链上,是一种新的胃癌相关抗原。     

抗人结肠癌单克隆抗体细胞株的建立及其免疫组化研究

本文报道了抗人结肠癌单克隆抗体3E_4细胞株的建立方法。其上清液与各种人癌细胞株及正常组织细胞等反应均阴性,仅与AFP似有弱交叉反应;经免疫组化研究,3E_4株单克隆抗体对结肠癌粘膜组织有一定选择性,但与癌旁正常结肠组织仍有一定交叉反应,属于抗结肠癌相关抗原的单克隆抗体。     

抗猪囊尾蚴KD26抗原单克隆抗体杂交瘤细胞株建立和特性鉴定

目的: 建立针对猪囊尾蚴KD26抗原的单克隆杂交瘤细胞株,生产相应的单克隆抗体,用于囊虫病免疫保护机制的基础研究和囊虫病临床免疫诊断的应用。方法: 用SDS-PAG E电泳,分离纯化制备猪囊尾蚴KD26抗原免疫6周龄BALB/C小鼠3次,无菌取出脾细胞与小鼠骨髓瘤细胞SP2/O(比率10:1),在50% PEG (4 000MW)作用下进行细胞融合,随后在HT培养液中进行选择性培养、筛选和克隆;常规的中期染色体法分析染色体,单克隆抗体鉴定采用免疫双扩散法测定其免疫球蛋白亚类,采用ELISA法测定单抗效价并作特异性鉴定。结果: 用常规的有限稀释法对杂交瘤细胞进行了3次筛选和克隆化后,获得了1株杂交瘤XH,染色体数为96,能稳定分泌抗猪囊尾蚴KD26单抗,单抗类型为IgG1,抗体滴度为1:1 000,其与血吸虫、弓形虫、细粒棘球绦虫抗原均不发生交叉反应。结论: 应用杂交瘤技术获得了1株能稳定分泌高滴度抗猪囊尾蚴KD26抗原的单克隆抗体细胞株。     

小鼠脾内免疫法制备重组弓形虫SAG1抗原单克隆抗体

 目的 制备小鼠抗重组弓形虫SAG1抗原单克隆抗体,用于早期弓形虫感染的抗原检测。方法 用弓形虫RH株重组抗原SAG1进行常规免疫结合小鼠脾内免疫,杂交瘤技术制备单克隆抗体。ELISA法筛选阳性克隆,经亚克隆建株。诱生腹水法制备抗体,protein-G亲合层析法进行纯化,测定其亚类、效价;Western blot法分析其特异性;夹心-ELISA法检测抗体的敏感性及特异性;检测弓形虫感染小鼠血液循环抗原,同时用PCR法检测弓形虫B1基因并比较结果。结果 获得2株抗重组弓形虫SAG1抗原单克隆抗体3B6、10C4,抗体均为IgG1,轻链均为κ链,Western blot显示2株单抗均能识别弓形虫天然的和重组的SAG1抗原。3B6、10C4敏感度分别为31.3 ng和62.5 ng,与血吸虫病、钩虫病及疟疾患者血清均无交叉反应。弓形虫早期感染检测PCR及ELISA法阳性检出率分别为63.2%、47.4%。结论 成功制备小鼠抗重组弓形虫SAG1抗原单克隆抗体,初步用于早期弓形虫感染诊断。     

鼠痘病毒单克隆抗体杂交瘤细胞株的建株及初步鉴定

目的:制备1株分泌高效价抗新分离的鼠疫病毒单克隆抗体的杂交瘤细胞,提高早期检测病毒的敏感性.方法:从可疑病鼠分离出的1株鼠痘病毒,并以此株制备抗鼠痘病毒为抗原,免疫BALB/C小鼠,取脾细胞与SP2/0骨髓瘤细胞融合.获1株稳定分泌抗鼠痘病毒(EV)单克隆抗体的杂交瘤细胞株(NB-4).结果:经初步鉴定证明,NB-4免疫球蛋白为IgG,针对的靶抗原分子质量为67ku,并对184份小鼠血清鼠痘病毒抗原进行检测.结论:NB-4有可能成为检测鼠痘病毒抗原较敏感的工具.     

胃癌单克隆抗体MG7相应抗原与DBA受体在胃癌细胞中的电镜双标记定位

作者应用两种胶体金探针,于胃癌组织冰冻超薄切片上,对胃癌单抗MG7相应抗原与DBA受体进行了电镜双标记定位.结果显示,二者在胃癌细胞中的分布基本一致,主要定位于细胞膜表面、细胞内微囊腔面及部分粘液泡内,但在同一区域内,MG7与相应抗原结合位点的数目明显多于DBA与相应受体结合位点.提示胃癌单抗MG7相应抗原的分布符合分泌性抗原的特点.表明胃癌单抗MG7与相应抗原有很强的识别和结合能力.本实验结果可为单克隆抗体应用于胃癌导向治疗提供超微形态学依据.     

MUCOUS CELL CARCINOMA OF THE STOMACHA HISTOCHEMICAL AND IMMUNOHISTOCHEMICAL STUDY OF 50 CASES

This article reports histochemical and immuno-histochemical studies on 50 cases of gastric mucouscell carcinoma. The purpose was to explore themucinous antigenicity of gastric cancer by monoclo.nal antibodies to small and large intestinal mucin antigens (SIMA and LIMA), and to find the relation-ship between mucinous antigens and sulphomucin,O-acety l(O-ac) sialomucin. The results demonstratethat 98% of stomach mucous cell carcinomas are ofmixed gastrointestinal type representing differentsecretory phases of the same cancer cells and therefore indicating that the features of m相似文献   

HUMAN COLONIC CANCER ASSOCIATED ANTIGENS DETECTED BY THREE MONOCLONAL ANTIBODIES

Three monoclonal antibodies (Hb3, Ab5, and Ad5) were produced with spleen cells from mice im munized against the human colon cancer cell line HT-29. Hb3 reacted with 86%, while Ab3 and Ad5 with 40-60%, of the colon cancer tissues tested. Re- activity was also seen with the cancerous stomach, pancreas, lung, nasopharynx, breast and prostate, although in Hb3 the intensity of immunofluorescent staining of these tissues was not as great as of colon cancer tissues. None of the monoclonal antibodies reacted with normal colon samples so they were able to distinguish the normal from the cancerous colon tissues. Yet monoclonal antibodies did react with the normal duodenum, stomach, pancreas, lung, and kidney. The three antibodies can be differentiated from each other on the basis of their cell and tissue specificities and from the colon cancer reactive mo- noclonal antibody 19-9. Neuraminidase digestion of the tissue or cells had no effect on their reactivities. - 63 -     

Purification and partial characterization of a new group of gastric cancer associated antigens.

The corresponding antigens of MG series monoclonal antibodies (MG5, MG9, MGd1 and MGe1) against gastric cancer were purified and partially characterized. Each of these monoclonal antibodies was purified by passing through a DEAE-52 cellulose columns and covalently coupled with CNBr-activated sepharose 4B successively. By means of concanavalin A and antibody affinity chromatography, the corresponding antigens of MG series McAb were extracted from gastric cancer tissue respectively. Immunological and biochemical studies confirmed that the corresponding antigens of MG series McAb were a new group of gastric cancer associated neutral glycolipid and glycoprotein antigens.

     

人类结肠癌单克隆抗体的研究

用大肠癌HT29作抗原免疫小鼠,取其脾脏的淋巴细胞与骨髓瘤细胞融合,产生三株杂交瘤细胞——Ga_3、Gb_6和Cb-5,其所作用的抗原为肿瘤相关抗原。间接免疫荧光的研究发现:三株抗体与肿瘤的石腊切片无反应,但与冰冻切片及大肠癌细胞株作短期固定者反应阳性。Ga_3属IgG_(2a),Gb_6和Cb_5属IgM。三株抗体可能有助于结肠癌的免疫诊断。     

MOUSE AND HUMAN MONOCLONAL ANTIBODIES AGAINSTGASTRIC CANCERPREPARATION AND CLINICAL APPLICATION

Ten clones of mouse and two clones, oF hu-man phybridoma cell producing monoclonal an- tibodies against gastric cancer were established. The nature, specificity and affinity of these mo- noclonal antibodies for gastric cancer associated antigens were demonstrated. They were used preliminarily in the diagnosis of gastric carcino- ma with encouraging results. Cultivated gastric cancer cells or tumor cells from surgically resected gastric cancer or affected lymph nodes were used to immunize Balb/c mice. The lymphocytes harvested from the spleen after immunization were fused with mouse myeloma cell line SP2/0. Ten kinds of mouse monoclonal antibodies reactive with different histopathologi- cal types of gastric carcinomas have been prepar~ ed. Two kinds of human monoclonal antibodies have also been produced by fusion of the lym- phocytes from the draining lymph nodes with mouse-human heteromyeloma cell line SHM-D33. Considerable amounts of McAbs can be obtained from the cell culture supernatant or from ascitic fluid of mice with intraperitoneally transplanted hybridoma cells.     

人肺腺癌细胞系SLC-89单克隆抗体的研制

用本室建立的“宣威”人肺腺癌细胞系SLC—89作抗原制备了三株抗人肺癌单克隆抗体细胞株4E_(12),4F_(12)和4H_2。它们在体外已培养了8个多月,多次液氮冻存后复苏,生长良好,并继续分泌抗体,经检测,所分泌的抗体对肺癌有较好的反应性,相应抗原在常规石蜡组织切片中仍较稳定,因此有可能作为临床病理诊断试剂。     

免疫酶技术检查EHF患者外周血单个核细胞中病毒结构蛋白抗原的研究

２２例ＥＨＦ患者于不同病期外周血单个核细胞涂片，用已知抗病毒核蛋白的ＭｃＡｂＡ３５及抗膜蛋白的ＭｃＡｂ３Ｇ＿１进行免疫酶染色，以检查病毒结构蛋白抗原。３病日时抗原检出率为６３．１％，７病日时达１００％，１７病日时仅为１８．２％，与抗体应答规律一致，只是其检出比抗体检出约提早３ｄ，且核、膜蛋白抗原的检出率间差异无统计学意义，结合ＥＨＦ患者对病毒结构蛋白免疫应答的特点，可预测患者的转归。同时该法也可用于ＥＨＰ患者早期快速诊断，并值得推广应用于其它病毒性疾病的诊断中。     

Strong antibody reaction against glycosphingolipids injected in liposome-embedded forms in beta3GN-T5 knockout mice

It is known that mutant mice of the beta-1,3-N-acetylglucosaminyltransferase gene (beta3Gn-T5) respond well to T-cell dependent and independent antigens. Here, we examined the effectiveness of anti-ganglioside antibody generation by immunization of beta3Gn-T5 mutant mice with liposome-embedded glycosphingolipids such as GD1a and GT1b. Consequently, the mutant mice showed a more efficient generation of anti-GD1a or anti-GT1b antibodies than wild-type mice in an enzyme-linked immunosorbent assay using sera during immunization. Thus, the beta3Gn-T5 deficient mutant mice proved more responsive than wild-type mice to not only protein antigens, but also to carbohydrates in glycolipids. Furthermore, about 50% of monoclonal antibodies generated using splenocytes of the immunized mutant mice were of the IgG class. Besides general high responsiveness to proteins and glycolipids, it could be expected that the mutant mice of beta3Gn-T5 would be useful in the generation of monoclonal antibodies towards lacto-/neolacto-series glycolipids, since these mutants lack lacto-/neolacto-series glycolipids. In fact, they showed a good serum response in immuno-fluorescence assay with cultured living cells when immunized by glycolipids extracted from ovarian cancer cell lines. These results suggested that beta3Gn-T5 mutant mice are useful for the generation of anti-glycolipid antigens with lacto-/neolacto-core structures expressed in cancer cells.     

肺鳞癌患者胸水免疫复合物抗原及抗体组份的免疫学特异性分析

本文利用Protein A-Sepharose CL-4B亲和层析法纯化肺鳞癌患者胸水免疫复合物(IC);并在解离的条件下(8mol/L尿素),用QAE-Sephadex A-50离子交换柱层析法分离胸水免疫复合物中的抗原、抗体组份。IC抗体组份经间接免疫荧光法鉴定具有相对肺癌的反应特异性;用IC抗原组份制备的特异性抗血清经正常人组织、血浆等免疫吸附剂充分吸附后,仍保留与原胸水IC中分离所得抗体组份相似的抗肿瘤相关抗原反应特异性。该法为研究肿瘤宿主体内免疫调节反应提供了较直接的途径。分离所得抗原组份制备的抗血清可作为研究、检测宿主体内肿瘤相关抗原的特异探针。