旨在建立注射用重组抗肿瘤坏死因子-α人鼠嵌合单克隆抗体病毒去除/灭活工艺,并进行效果验证。膜过滤工艺去除病毒验证结果表明,膜过滤后的蛋白质回收率在98%以上,分子排阻色谱纯度在膜过滤前后没有明显变化;经测试,膜过滤后小鼠白血病...旨在建立注射用重组抗肿瘤坏死因子-α人鼠嵌合单克隆抗体病毒去除/灭活工艺,并进行效果验证。膜过滤工艺去除病毒验证结果表明,膜过滤后的蛋白质回收率在98%以上,分子排阻色谱纯度在膜过滤前后没有明显变化;经测试,膜过滤后小鼠白血病病毒、小鼠微小病毒、呼肠孤病毒滴度的降幅均大于4 lg TCID 50/0.1 mL。低pH值孵放灭活病毒验证工艺结果表明,低pH值孵放前后蛋白质含量、分子排阻色谱纯度没有明显变化;测试结果表明,室温处理时间≥0.5 h,小鼠白血病病毒、伪狂犬病毒的滴度降幅均大于4 lg TCID 50/0.1 mL。该去除/灭活效果均符合《生物组织提取制品和真核细胞表达制品的病毒安全性评价技术审评一般原则》的要求,建立的工艺有效保证了产品的质量及安全性。展开更多
Background: Rituximab is an anti-CD20 chimeric antibody that selectively targets B lymphocytes. Recently, it has been reported to be beneficial in treating pemphigus vulgaris. Objective: Our aim was to review the Engl...Background: Rituximab is an anti-CD20 chimeric antibody that selectively targets B lymphocytes. Recently, it has been reported to be beneficial in treating pemphigus vulgaris. Objective: Our aim was to review the English-language literature on the treatment of pemphigus vulgaris (PV) with rituximab and to determine its efficacy and influence on clinical outcome(s). Material and methods: A retrospective review of the literature on the use of rituximab in the treatment of PV was conducted. Seventeen patients in 10 reports were described and their data were reviewed. Results: The majority of patients received one course of rituximab along with conventional immunosuppressive therapy as concomitant therapy; 88%of the patients demonstrated improvement. More than half of the patients were followed up for more than 6 months after rituximab treatment; they appeared to be clinically disease free, but were still receiving conventional immunosuppressive therapy. Side effects in most patients were transient and infusion related. Serious infections occurred in 4 patients. One patient died. Limitations: The sample size of this study is small; there is no uniformity of data collection or measurement of key and critical indices, and follow-up was limited. Conclusion: Rituximab may be a promising agent in treatment of PV.展开更多
Background:Primary cutaneous B-cell lymphomas (PCBCLs)are characterized by restriction to the skin and a variable but mostly favourable prognosis. Since 1997 the recombinant, chimeric anti-CD20 antibody rituximab has ...Background:Primary cutaneous B-cell lymphomas (PCBCLs)are characterized by restriction to the skin and a variable but mostly favourable prognosis. Since 1997 the recombinant, chimeric anti-CD20 antibody rituximab has been used in patients suffering from non-Hodgkin,s B-cell lymphomas. Different studies have shown that the effectiveness and safety in the treatment of patients with low-grade follicular lymphoma is comparable to or even higher than the standard CHOP chemotherapy. So far it has been unclear whether an extended duration of therapy lead s to a benefit for the patients with PCBCL. Objectives:To evaluate the objective response rate, time to progression, remission quality and histological changes and to compare our data with the literature. Patients/Methods:Ten patients with PCBCL [eight with follicle centre cell lymphoma (FCCL), one with marginal zone lymphoma (MZL) and one with diffuse large B-cell lymphoma of the leg (DLBCL)]were treated by intravenous application of a chimeric antibody against the CD20 transmembrane antigen (rituximab) with a dosage of eight cycles, 375 mg m-2 body surface, weekly. Results:The treatment regimen resulted in clinical overall response in 9 of 10 patients, in particular there were seven complete responses (70%) plus two partial responses (20%). The median duration of remission (durable remission, DR) is 23 months (4-30 months) to date. Histological assessment of responses in four patients showed no tumour-specific infiltration. In two patients histology revealed a residual infiltration and in one patient an increasing infiltration. In two patients no histology was taken after treatment; one patient developed a new lesion. No severe side-effects occurred. Observed side-effects were two bacterial infections, two patients with shivering during infusion, one patient with sweating for months and one patient with persisting itching. As expected the B-cell count in peripheral blood was depressed in all patients after infusion. Conclusions:Intravenous therapy with eight cycles of the antiCD20 antibody rituximab is a non-toxic and effective treatment for a sub set of patients with PCBCL(relapsed,aggressiveentity, old patients, multiple lesions) with a long DR.展开更多
利用淋巴细胞杂交瘤技术建立分泌特异性McAb杂交瘤细胞系,从中提取已重排的编码功能性Ig DNA 基因组(组建DNA文库)或成熟的IgH,L链mRNA片段(组建cDNA文库及制备探针),从而获得编码特异性McAb的V区基因。利用基因重组技术,将此来源于小...利用淋巴细胞杂交瘤技术建立分泌特异性McAb杂交瘤细胞系,从中提取已重排的编码功能性Ig DNA 基因组(组建DNA文库)或成熟的IgH,L链mRNA片段(组建cDNA文库及制备探针),从而获得编码特异性McAb的V区基因。利用基因重组技术,将此来源于小鼠特异性IgH、L链的V区基因分别与编码人IgH、L链的C区基因相重组,构成鼠/人嵌合的Ig重组基因。然后将此重组嵌合Ig基因导入真核表达质粒(多为pSV2-ypt和pSV2-neo)_4经磷酸钙沉淀技术,原生质体融合技术或电穿孔导入技术(以后2种方法较为实用且转化频率较高),将此质粒转染哺乳动物非分泌型骨髓瘤细胞如Sp2/0等,经选择性培养基培养,从中筛选分必完整功能性的鼠/人嵌合McAb。将此cMcAb临床应用于人体内进行诊断和治疗时,具有原鼠亲本McAb特异性结合抗原的能力,同时还具备优于亲本鼠McAb的介导补体,细胞对靶抗原的杀伤和吞噬作用。而且不引起过敏反应或干扰治疗效果。从而给临床诊断,治疗恶性肿瘤。病毒,细菌等感染提供了一种新颖的免疫治疗试剂、本文对近年来鼠/人嵌合McAb的研究进展,基本技术及原理,优越性及不足之处作了简要阐述,并认为制备嵌合McAb是今后制备并取代人McAb研制及应用的一种发展趋向。展开更多
文摘旨在建立注射用重组抗肿瘤坏死因子-α人鼠嵌合单克隆抗体病毒去除/灭活工艺,并进行效果验证。膜过滤工艺去除病毒验证结果表明,膜过滤后的蛋白质回收率在98%以上,分子排阻色谱纯度在膜过滤前后没有明显变化;经测试,膜过滤后小鼠白血病病毒、小鼠微小病毒、呼肠孤病毒滴度的降幅均大于4 lg TCID 50/0.1 mL。低pH值孵放灭活病毒验证工艺结果表明,低pH值孵放前后蛋白质含量、分子排阻色谱纯度没有明显变化;测试结果表明,室温处理时间≥0.5 h,小鼠白血病病毒、伪狂犬病毒的滴度降幅均大于4 lg TCID 50/0.1 mL。该去除/灭活效果均符合《生物组织提取制品和真核细胞表达制品的病毒安全性评价技术审评一般原则》的要求,建立的工艺有效保证了产品的质量及安全性。
文摘Background: Rituximab is an anti-CD20 chimeric antibody that selectively targets B lymphocytes. Recently, it has been reported to be beneficial in treating pemphigus vulgaris. Objective: Our aim was to review the English-language literature on the treatment of pemphigus vulgaris (PV) with rituximab and to determine its efficacy and influence on clinical outcome(s). Material and methods: A retrospective review of the literature on the use of rituximab in the treatment of PV was conducted. Seventeen patients in 10 reports were described and their data were reviewed. Results: The majority of patients received one course of rituximab along with conventional immunosuppressive therapy as concomitant therapy; 88%of the patients demonstrated improvement. More than half of the patients were followed up for more than 6 months after rituximab treatment; they appeared to be clinically disease free, but were still receiving conventional immunosuppressive therapy. Side effects in most patients were transient and infusion related. Serious infections occurred in 4 patients. One patient died. Limitations: The sample size of this study is small; there is no uniformity of data collection or measurement of key and critical indices, and follow-up was limited. Conclusion: Rituximab may be a promising agent in treatment of PV.
文摘Background:Primary cutaneous B-cell lymphomas (PCBCLs)are characterized by restriction to the skin and a variable but mostly favourable prognosis. Since 1997 the recombinant, chimeric anti-CD20 antibody rituximab has been used in patients suffering from non-Hodgkin,s B-cell lymphomas. Different studies have shown that the effectiveness and safety in the treatment of patients with low-grade follicular lymphoma is comparable to or even higher than the standard CHOP chemotherapy. So far it has been unclear whether an extended duration of therapy lead s to a benefit for the patients with PCBCL. Objectives:To evaluate the objective response rate, time to progression, remission quality and histological changes and to compare our data with the literature. Patients/Methods:Ten patients with PCBCL [eight with follicle centre cell lymphoma (FCCL), one with marginal zone lymphoma (MZL) and one with diffuse large B-cell lymphoma of the leg (DLBCL)]were treated by intravenous application of a chimeric antibody against the CD20 transmembrane antigen (rituximab) with a dosage of eight cycles, 375 mg m-2 body surface, weekly. Results:The treatment regimen resulted in clinical overall response in 9 of 10 patients, in particular there were seven complete responses (70%) plus two partial responses (20%). The median duration of remission (durable remission, DR) is 23 months (4-30 months) to date. Histological assessment of responses in four patients showed no tumour-specific infiltration. In two patients histology revealed a residual infiltration and in one patient an increasing infiltration. In two patients no histology was taken after treatment; one patient developed a new lesion. No severe side-effects occurred. Observed side-effects were two bacterial infections, two patients with shivering during infusion, one patient with sweating for months and one patient with persisting itching. As expected the B-cell count in peripheral blood was depressed in all patients after infusion. Conclusions:Intravenous therapy with eight cycles of the antiCD20 antibody rituximab is a non-toxic and effective treatment for a sub set of patients with PCBCL(relapsed,aggressiveentity, old patients, multiple lesions) with a long DR.
文摘利用淋巴细胞杂交瘤技术建立分泌特异性McAb杂交瘤细胞系,从中提取已重排的编码功能性Ig DNA 基因组(组建DNA文库)或成熟的IgH,L链mRNA片段(组建cDNA文库及制备探针),从而获得编码特异性McAb的V区基因。利用基因重组技术,将此来源于小鼠特异性IgH、L链的V区基因分别与编码人IgH、L链的C区基因相重组,构成鼠/人嵌合的Ig重组基因。然后将此重组嵌合Ig基因导入真核表达质粒(多为pSV2-ypt和pSV2-neo)_4经磷酸钙沉淀技术,原生质体融合技术或电穿孔导入技术(以后2种方法较为实用且转化频率较高),将此质粒转染哺乳动物非分泌型骨髓瘤细胞如Sp2/0等,经选择性培养基培养,从中筛选分必完整功能性的鼠/人嵌合McAb。将此cMcAb临床应用于人体内进行诊断和治疗时,具有原鼠亲本McAb特异性结合抗原的能力,同时还具备优于亲本鼠McAb的介导补体,细胞对靶抗原的杀伤和吞噬作用。而且不引起过敏反应或干扰治疗效果。从而给临床诊断,治疗恶性肿瘤。病毒,细菌等感染提供了一种新颖的免疫治疗试剂、本文对近年来鼠/人嵌合McAb的研究进展,基本技术及原理,优越性及不足之处作了简要阐述,并认为制备嵌合McAb是今后制备并取代人McAb研制及应用的一种发展趋向。
