共查询到17条相似文献,搜索用时 281 毫秒
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用人血型糖蛋白 A 和人红细胞分别免疫 BALB/C 小鼠。用被免疫的 BALB/C 小鼠脾细胞与 SP2/0骨髓瘤细胞融合,在37℃、5%二氧化碳培养箱中孵育10~15天,然后用 OM 和 ON 型指示红细胞分别检测培养上清液的凝集情况,筛选出能分泌高特异性和高效价抗 M 和抗 N 抗体的细胞株,并建立了 GM_4H3、GM_4H_4、N_2A_3和 N_2D_(10)细胞株。这些细胞株可持续分泌免疫球蛋白 G 类抗 M、抗 N 抗体。应用这些抗体,通过血凝法、解离法和 ELISA 斑点法,可进行血及血痕的 MN 分型。在血型检验中,优于多克隆抗 M、抗N 血清。 相似文献
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抗丁丙诺啡单克隆抗体的制备 总被引:2,自引:2,他引:0
目的建立抗丁丙诺啡单克隆抗体的杂交瘤细胞株,制备高特异性的丁丙诺啡单克隆抗体,并对其免疫学特性进行鉴定。方法在丁丙诺啡的分子上连接活性羧基基团,通过缩合反应将丁丙诺啡半抗原连接于血蓝蛋白(KLH)和小牛血清白蛋白(BSA),形成完全抗原。以完全抗原免疫Balb/c小鼠,通过细胞融合,筛选等杂交瘤技术,建立稳定的分泌抗丁丙诺啡单克隆抗体的杂交瘤细胞株。通过腹腔注射杂交瘤细胞,诱导小鼠产生含有单抗的腹水。用辛酸-硫酸铵加亲和层析法纯化抗丁丙诺啡单克隆抗体。采用酶联免疫反应和胶体金膜层析实验测定丁丙诺啡单抗的特异性以及免疫反应动力学参数。结果共获得3株分泌抗丁丙诺啡单克隆抗体的杂交瘤细胞株,分别命名为7E6,6G4和3C2。7E6,6G4抗体灵敏度为10.0ng/ml,3C2抗体灵敏度为20.0ng/ml。7E6,6CA和3C2抗体的亲和常数分别为3.6×10^-9 mol/L,4.3×10^-9 mol/L和6.3×10^-9 mol/L。特异性测试结果表明7E6和6G4抗体与40种药物、毒品无任何交叉反应,而3C2抗体与吗啡有交叉反应。结论杂交瘤细胞株7E6和6G4产生的抗丁丙诺啡单克隆抗体具有很高的特异性和灵敏度。 相似文献
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分泌抗人精液特异蛋白P_(30)单克隆抗体杂交瘤细胞系的建立和初步应用 总被引:1,自引:0,他引:1
作者通过杂交瘤技术建立了9株产生抗精浆特异蛋白 P_(30) 单克隆抗体的杂交瘤细胞系。它们是由 SP2/0骨髓瘤与经 P_(30) 免疫的 BALB/C 小鼠脾细胞按常规方法进行细胞融合、并经克隆化筛选得出.这些细胞株均经体外培养3个月以上能够稳定分泌抗 P_(30) 单克隆抗体。该抗体只能识别纯化的 P_(30) 和精液中的 P_(30) ;与人精液以外的其他体液和多种人体组织无交叉反应;与几种常见动物的精液和血液无交叉反应。这些 P_(30) 单克隆抗体均属 IgG 类和 IgG_1亚类。其培养上清液和腹水的抗体效价最高分别达到320和128,000。以 ELISA 法应用这些单克隆抗体能很好地鉴定精液和精斑。 相似文献
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分泌抗人精子单克隆抗体杂交瘤细胞株的建立及鉴定 总被引:1,自引:0,他引:1
建立两株分泌抗人精子单克隆抗体的杂交瘤细胞株G_8与E_(10),采用洗涤人精子免疫Balb/c小鼠,取免疫脾与Balb/c小鼠的Sp2/0骨髓瘤细胞融合,杂交瘤细胞诱生腹水的方法。Elisa,IIF或酶标抗体免疫组化法结果发现,G_8与E_(10)单抗与人精子(射出精子、睾丸、副睾管及输精管中的精子)及睾丸各级生殖细胞均发生反应;与人甲状腺上皮细胞、肾小管上皮细胞、胰腺导管上皮细胞以及胰岛均发生不同程度的交叉反应,与其他33种人体正常组织、9种人体液与分泌液、8种不同种族的动物精子均不发生交叉反应,说明在一般情况下这两种抗体可用于鉴定人类的精液斑以及精液与阴道分泌液的混合斑。两种单抗与人精子的顶体部、赤道部、后核帽、颈部及中段结合,证明精子特异性表面抗原分布于这几个区域。两种抗体均属IgG_1,G_8与E_(10)抗体的效价分别为1:512与1:1024。 相似文献
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The localization of A and B antigens in the organs of blood group AB individuals has been studied using a double immunoenzymatic labeling method. Both A and B antigens were found in the various epithelial cells of these organs, but the epithelial cells could be classified into the following four types depending on the reaction pattern with anti-A and anti-B sera: type 1: cells that stained positive with both anti-A and anti-B sera; type 2: cells that stained positive with anti-A serum only; type 3: cells that stained positive with anti-B serum only; type 4: cells that were negative with both sera. The distribution of each of these epithelial cell types varied considerably, even in the same tissue and individual. Our results seem to suggest that a dissociation in the conversion to the A and B antigens occurs in the tissue of individuals belonging to blood group AB and that the degree of this dissociation varies from tissue to tissue and from cell to cell. 相似文献
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Kolokolova GP 《Sudebno-meditsinskaia ekspertiza》2006,49(4):32-34
Experimental spots of the saliva, sperm, vaginal secretion from persons with groups Ase, Bse, ABSe and Abse were studied with mixed agglutination reaction (MAR) using hemagglutinating sera anti-A and anti-B (heteroimmune, isoimmune), monoclonal antibodies. MAR with monoclonal antibodies was able to diagnose ABSe group not only in the spots of saliva, but also in the spots of sperm and vaginal secretion where heteroimmune sera failed. 相似文献
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目的建立分泌抗三唑仑代谢物α-羟基三唑仑单克隆抗体的杂交瘤细胞株,制备高特异性的三唑仑代谢物单克隆抗体,为三唑仑及其代谢物免疫分析方法的开发奠定基础。方法在三唑仑分子的6位苯环对位上引入活性氨基基团,然后通过缩合反应分别与匙孔血蓝蛋白(KLH)和牛血清白蛋白(BSA)相偶联形成完全抗原。以三唑仑-KLH免疫Balb/c小鼠,通过细胞融合,筛选等杂交瘤技术建立稳定的分泌特异性单克隆抗体的杂交瘤细胞株。纯化后的单克隆抗体,分别用SDS-PAGE电泳法、间接ELISA法和胶体金免疫层析法对其纯度、效价及灵敏度和特异性进行测定。结果获得3株能稳定分泌三唑仑代谢物单克隆抗体的杂交瘤细胞株,分别命名为2G4,4B2和5H6。2G4和4B2抗体只与三唑仑代谢物α-羟基三唑仑有反应,灵敏度分别为500ng/mL和750ng/mL。与其他参试物无交叉反应。因5H6抗体为IgM,考虑到纯化难度和实际应用的限制,暂未做深入研究。结论本研究制备的2G4和4B2单克隆抗体仅识别三唑仑代谢物α-羟基三唑仑,具有高度特异性和灵敏度。 相似文献
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The unlabeled antibody (PAP) immunoperoxidase technique was applied to the ABO blood grouping of human scalp hairs. Hair samples were subjected to longitudinal- or cross-sectioning, thus obtaining suitable samples for subsequent immunostaining. The immunostaining was carried out using rabbit anti-A and anti-B sera as the primary antibodies. With this technique, the group-specific staining which is revealed as a dark brown precipitate was clearly observed within the medullae of the hair shaft, and depending on the presence or absence of these precipitates, respective blood groups of unknown hair samples were determined. At the hair root, on the other hand, positive stainings were observed not only in medullary cells but also in some cortical cells of the keratogenous zone. From the present study, it can be safely said that this technique is of practical use for the ABO blood grouping from a minute (less than 3 mm) hair sample. 相似文献
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A simple method for purification of anti-A and anti-B antibodies using glutaraldehyde-fixed human erythrocytes is described. Specific antibodies were first absorbed with the corresponding cells, then eluted by heating at 53 degrees-55 degrees C for 15 min. The method is simple and highly efficient with a fair recovery of 15.6%-34.4%. 相似文献
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巴比妥单克隆抗体制备及其免疫学特性鉴定 总被引:1,自引:1,他引:0
目的建立抗巴比妥单克隆抗体杂交瘤细胞株,制备抗巴比妥单克隆抗体,并对其免疫学特性进行鉴定。方法将巴比妥分子环状丙二酰脲环上氨基引入活性羧基,然后通过缩合反应分别与匙孔血蓝蛋白(KLH)和牛血清白蛋白(BSA)偶联,得到完全抗原BAR-KLH和BAR-BSA,并用紫外分光光度计鉴定。用BAR-KLH免疫Balb/C小鼠,利用细胞融合技术建立稳定分泌抗巴比妥单克隆抗体的杂交瘤细胞株;采用体内诱生腹水法制备巴比妥单克隆抗体,饱和硫酸铵法、亲和层析法纯化,并进行免疫学特性鉴定。结果完全抗原偶联成功,其偶联率为30∶1;筛选出1株杂交瘤细胞命名为5C6,其产生的单抗效价为1∶6.4×104,属于IgG1/kappa,抗体亲和力常数为2.27×108L/moL。纯化后的巴比妥单克隆抗体用ELISA法检测其灵敏度为130ng/mL,并与其他7种镇静催眠类药物交叉反应率小于1%。结论本研究制备的杂交瘤细胞株5C6分泌的抗体具有高特异性和灵敏度。 相似文献