以纯化的经鸡胚成纤维细胞（CEF）培养的马立克病病毒（MDV）超强毒株GX0101病毒粒子作为免疫原，制备特异性识别MDV的单克隆抗体。GX0101感染CEF后第96小时收集细胞并反复冻融3次，离心取上清，选用100ku的切向流超滤离心管浓缩，浓缩...以纯化的经鸡胚成纤维细胞（CEF）培养的马立克病病毒（MDV）超强毒株GX0101病毒粒子作为免疫原，制备特异性识别MDV的单克隆抗体。GX0101感染CEF后第96小时收集细胞并反复冻融3次，离心取上清，选用100ku的切向流超滤离心管浓缩，浓缩液通过Sepharose 4 Fast Flow（4FF）凝胶作为层析介质分离纯化MDV病毒粒子，聚合酶链反应（PCR）和实时荧光定量PCR分析不同收集峰MDv病毒含量，并经过透射电子显微镜观察到直径约100nm的MDV病毒粒子。进一步用100ku的切向流超滤离心管浓缩,制备峰尖病毒浓缩液，以此免疫雌性BALB／c小鼠制备单克隆抗体。通过免疫过氧化物酶单层细胞试验（IPMA）和Western—blot筛选,最终获得107个阳性克隆。由其中1株强阳性单克隆抗体杂交瘤细胞株1082制备的单克隆抗体腹水1PMA效价为1：2．56×10^6。本研究结果为后续MDV单克隆抗体的筛选、鉴定及诊断试剂研发奠定了重要基础。展开更多
Three anti-EGF receptor MoAbs were used in these studies. Administration of MoAbs 3 and 176 inhibited tumor formation in nude mice by CNE-2, a poorly differentiated nasopharyngeal carcinoma cell line and A431, an epid...Three anti-EGF receptor MoAbs were used in these studies. Administration of MoAbs 3 and 176 inhibited tumor formation in nude mice by CNE-2, a poorly differentiated nasopharyngeal carcinoma cell line and A431, an epidermoid carcinoma cell line. When the same MoAbs were used in treatment against HeLa, a cervical carcinoma, tumor growth was not affected. The number of EGF receptors and apparent dissociation constants for ^(125)I-EGF on CNE-2 and A431 was 1.3×10^(?)/cell (Kd 7.7×10^(-8)mol/L) and 1.4×10~6/cell(Kd 2.4×10^(-9)mol/L), respectively. Both MoAbs 3 and 176, capable of competing with EGF for receptor binding, showed significant tumor growth inhibition. MoAb 101 was incapable of blocking the binding of EGF to its receptor, and not as effective as MoAbs 3 and 176 in tumor growth inhibition. Our observation is that the MoAb anti-EGF receptor is cytostatic rather than cytocidal, in vitro against CNE-2 and A431.展开更多
文摘以纯化的经鸡胚成纤维细胞（CEF）培养的马立克病病毒（MDV）超强毒株GX0101病毒粒子作为免疫原，制备特异性识别MDV的单克隆抗体。GX0101感染CEF后第96小时收集细胞并反复冻融3次，离心取上清，选用100ku的切向流超滤离心管浓缩，浓缩液通过Sepharose 4 Fast Flow（4FF）凝胶作为层析介质分离纯化MDV病毒粒子，聚合酶链反应（PCR）和实时荧光定量PCR分析不同收集峰MDv病毒含量，并经过透射电子显微镜观察到直径约100nm的MDV病毒粒子。进一步用100ku的切向流超滤离心管浓缩,制备峰尖病毒浓缩液，以此免疫雌性BALB／c小鼠制备单克隆抗体。通过免疫过氧化物酶单层细胞试验（IPMA）和Western—blot筛选,最终获得107个阳性克隆。由其中1株强阳性单克隆抗体杂交瘤细胞株1082制备的单克隆抗体腹水1PMA效价为1：2．56×10^6。本研究结果为后续MDV单克隆抗体的筛选、鉴定及诊断试剂研发奠定了重要基础。
文摘Three anti-EGF receptor MoAbs were used in these studies. Administration of MoAbs 3 and 176 inhibited tumor formation in nude mice by CNE-2, a poorly differentiated nasopharyngeal carcinoma cell line and A431, an epidermoid carcinoma cell line. When the same MoAbs were used in treatment against HeLa, a cervical carcinoma, tumor growth was not affected. The number of EGF receptors and apparent dissociation constants for ^(125)I-EGF on CNE-2 and A431 was 1.3×10^(?)/cell (Kd 7.7×10^(-8)mol/L) and 1.4×10~6/cell(Kd 2.4×10^(-9)mol/L), respectively. Both MoAbs 3 and 176, capable of competing with EGF for receptor binding, showed significant tumor growth inhibition. MoAb 101 was incapable of blocking the binding of EGF to its receptor, and not as effective as MoAbs 3 and 176 in tumor growth inhibition. Our observation is that the MoAb anti-EGF receptor is cytostatic rather than cytocidal, in vitro against CNE-2 and A431.
