Anti-HER2 monoclonal antibody （Sc7301）-paclitaxel （TAX） immunoconjugate was pre- pared and its specific binding to tumor cells was investigated in this study. Sc7301 was conjugated to TAX by the active ester metho...Anti-HER2 monoclonal antibody （Sc7301）-paclitaxel （TAX） immunoconjugate was pre- pared and its specific binding to tumor cells was investigated in this study. Sc7301 was conjugated to TAX by the active ester method and then the TAX-Sc7301 immunoconjugate was obtained. After purification and labeling by Cyano-fluorescein isothiocyanate （FITC）, the specific binding of TAX-Sc7301 to HER2-positive tumor cells （SKOV3） and HER2-negative tumor cells （HepG2） was evaluated respectively. TAX-Sc7301 （20 nmol/L） showed distinct specific binding to SKOV3 cells rather than HepG2 cells. And the uptake of the immunoconjugate by SKOV3 cells was increased with the TAX-So7301 concentration （3-48 nmol/L） and the incubation time （P〈0.05）. It was concluded that the TAX-Sc7301 immunoconjugate is ootentially applicable as a targeted agent against HER2-10ositive tumor cells.展开更多
AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 ...AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 in liver tissue of patients with hepatic cirrhosis, and the correlation between TIMPs in liver and those in sera so as to discuss whether TIMPs can be used as a diagnosis index of hepatic fibrosis. METHODS: The monoclonal antibodies (McAbs) of TIMP-1 and TIMP-2 were used to sensitize erythrocytes, and solid-phase absorption to sensitized erythrocytes (SPASE) was used to detect TIMP-1 and TIMP-2 in the sera of patients with hepatic cirrhosis. Meanwhile, with the method of in situ hybridization and immunohistochemistry, we studied the mRNA expression and antigen location of TIMP-1 and TIMP-2 in the livers of 40 hepatic cirrhosis patients with pathologic diagnosis. RESULTS: With SPASE, they were 16.4% higher in the acute hepatitis group, 33.3% higher in the chronic hepatitis group, and the positive rates were 73.6% and 61.2% respectively in sera of hepatic cirrhosis patients, which were remarkably higher than those in chronic hepatitis and acute hepatitis group (P【0.001). In 40 samples of hepatic cirrhosis tissues, all of them showed positive expression of TIMP-1 and TIMP-2 mRNA detected with immunohistochemistry or in situ hybridization (positive rate was 100%). Expression of TIMPs in different degrees could be found in liver tissue with cirrhosis. TIMPs were located in cytoplasm of liver cells of patients with hepatic cirrhosis. There was a significant correlation between serum TIMPs level and liver TIMPs level. CONCLUSION: SPASE is a useful method to detect the TIMP-1 and TIMP-2 in sera of patients with hepatic cirrhosis, and TIMP-1 and TIMP-2 can be considered as a useful diagnostic index of hepatic fibrosis, especially TIMP-1.展开更多
目的观察重组全人源抗新型冠状病毒单克隆抗体(2B11)注射液对恒河猴2周重复给药的毒性反应,确定无毒性反应的安全剂量及给药剂量、给药时间与毒性之间的关系,为临床用药提供参考。方法30只健康恒河猴,随机分成3组,每组10只,雌雄各半。...目的观察重组全人源抗新型冠状病毒单克隆抗体(2B11)注射液对恒河猴2周重复给药的毒性反应,确定无毒性反应的安全剂量及给药剂量、给药时间与毒性之间的关系,为临床用药提供参考。方法30只健康恒河猴,随机分成3组,每组10只,雌雄各半。溶媒对照组给予0.9%氯化钠注射液,实验组分别给予2B11100和400 mg·kg^(-1)。每隔6 d iv给药1次,2周共给药3次,停药后恢复9周。实验期间进行一般症状观察、体重、摄食量、体温、眼科检查、血压、心电图、血常规、止凝血、血液生化及电解质、尿液、系统解剖、脏器系数、组织病理学和免疫学检测,同时进行抗药抗体(ADA)及血药浓度检测,并分析毒代动力学参数。结果实验期间2B112个剂量组恒河猴一般症状、体重、摄食量、体温、眼科检查、血压、心电图、血常规、止凝血、血液生化及电解质、尿液、脏器系数、组织病理学和免疫学检测等指标均未见与受试物有关的明显改变。2B112个剂量组均未检测到ADA,血浆药物浓度变化基本一致,且与给药剂量呈正比,峰浓度之比和暴露量之比也与给药剂量呈正比,2B11注射液在体内基本呈线性动力学特征。结论在本实验条件下,恒河猴2周重复iv给予2B113次是安全的,400 mg·kg^(-1)未观察到临床不良反应。展开更多
Human epidermal growth factor receptor 2(HER2)is an important biomarker for detection and treatment of breast cancer.In this study,we developed monoclonal antibodies against the extracellular domain(ECD)of HER2 and es...Human epidermal growth factor receptor 2(HER2)is an important biomarker for detection and treatment of breast cancer.In this study,we developed monoclonal antibodies against the extracellular domain(ECD)of HER2 and established a rapid and accurate lateral flow immunoassay(LFIA)for use in community medical institutions.The gene sequence of human HER2-ECD was obtained from the National Center for Biotechnology Information(NCBI)to construct the expression plasmid.HER2-ECD protein expressed in HEK293F cells was used to immunize BALB/c mice.The monoclonal antibodies were produced in mouse ascites and isolated by hybridoma cell screening.Antibodies were analyzed for purity by SDS-PAGE(sodium dodecyl sulphate-polyacrylamide gel-electrophoresis)and affinity was assessed by enzyme-linked immunosorbent assay(ELISA)while subtypes were detected using the commercial kits.The HER2-ECD test strip was prepared based on the sandwich method and evaluated using a portable detection instrument.The affinity of the paired antibodies,4D8 and 8D9,both reached 1×108 L/mol.Both antibodies specifically recognized the HER2-ECD protein in serum.The limit of detection(LOD)of the gold nanoparticle(AuNP)-based LFIA was 1.7 ng/mL with a detection range of 1.7-400 ng/mL,and the performance of the HER2-ECD strip correlated well with that of a Siemens chemiluminescent immunoassay(CLIA)kit.In conclusion,the paired antibodies were successfully prepared with high affinity and specificity.The AuNP-based LFIA of HER2-ECD provides a fast and accurate method to detect the concentration of HER2-ECD in serum samples for clinical use in community medical institutions,and could contribute to determining the progress of the disease or the effectiveness of treatment.展开更多
基金supported by a grant from the National Natural Science Foundation of China(No.30300429)
文摘Anti-HER2 monoclonal antibody （Sc7301）-paclitaxel （TAX） immunoconjugate was pre- pared and its specific binding to tumor cells was investigated in this study. Sc7301 was conjugated to TAX by the active ester method and then the TAX-Sc7301 immunoconjugate was obtained. After purification and labeling by Cyano-fluorescein isothiocyanate （FITC）, the specific binding of TAX-Sc7301 to HER2-positive tumor cells （SKOV3） and HER2-negative tumor cells （HepG2） was evaluated respectively. TAX-Sc7301 （20 nmol/L） showed distinct specific binding to SKOV3 cells rather than HepG2 cells. And the uptake of the immunoconjugate by SKOV3 cells was increased with the TAX-So7301 concentration （3-48 nmol/L） and the incubation time （P〈0.05）. It was concluded that the TAX-Sc7301 immunoconjugate is ootentially applicable as a targeted agent against HER2-10ositive tumor cells.
基金the Postdoctoral Science Foundation of China,No.1999-10 State Postdoctoral Foundation Commission
文摘AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 in liver tissue of patients with hepatic cirrhosis, and the correlation between TIMPs in liver and those in sera so as to discuss whether TIMPs can be used as a diagnosis index of hepatic fibrosis. METHODS: The monoclonal antibodies (McAbs) of TIMP-1 and TIMP-2 were used to sensitize erythrocytes, and solid-phase absorption to sensitized erythrocytes (SPASE) was used to detect TIMP-1 and TIMP-2 in the sera of patients with hepatic cirrhosis. Meanwhile, with the method of in situ hybridization and immunohistochemistry, we studied the mRNA expression and antigen location of TIMP-1 and TIMP-2 in the livers of 40 hepatic cirrhosis patients with pathologic diagnosis. RESULTS: With SPASE, they were 16.4% higher in the acute hepatitis group, 33.3% higher in the chronic hepatitis group, and the positive rates were 73.6% and 61.2% respectively in sera of hepatic cirrhosis patients, which were remarkably higher than those in chronic hepatitis and acute hepatitis group (P【0.001). In 40 samples of hepatic cirrhosis tissues, all of them showed positive expression of TIMP-1 and TIMP-2 mRNA detected with immunohistochemistry or in situ hybridization (positive rate was 100%). Expression of TIMPs in different degrees could be found in liver tissue with cirrhosis. TIMPs were located in cytoplasm of liver cells of patients with hepatic cirrhosis. There was a significant correlation between serum TIMPs level and liver TIMPs level. CONCLUSION: SPASE is a useful method to detect the TIMP-1 and TIMP-2 in sera of patients with hepatic cirrhosis, and TIMP-1 and TIMP-2 can be considered as a useful diagnostic index of hepatic fibrosis, especially TIMP-1.
文摘目的观察重组全人源抗新型冠状病毒单克隆抗体(2B11)注射液对恒河猴2周重复给药的毒性反应,确定无毒性反应的安全剂量及给药剂量、给药时间与毒性之间的关系,为临床用药提供参考。方法30只健康恒河猴,随机分成3组,每组10只,雌雄各半。溶媒对照组给予0.9%氯化钠注射液,实验组分别给予2B11100和400 mg·kg^(-1)。每隔6 d iv给药1次,2周共给药3次,停药后恢复9周。实验期间进行一般症状观察、体重、摄食量、体温、眼科检查、血压、心电图、血常规、止凝血、血液生化及电解质、尿液、系统解剖、脏器系数、组织病理学和免疫学检测,同时进行抗药抗体(ADA)及血药浓度检测,并分析毒代动力学参数。结果实验期间2B112个剂量组恒河猴一般症状、体重、摄食量、体温、眼科检查、血压、心电图、血常规、止凝血、血液生化及电解质、尿液、脏器系数、组织病理学和免疫学检测等指标均未见与受试物有关的明显改变。2B112个剂量组均未检测到ADA,血浆药物浓度变化基本一致,且与给药剂量呈正比,峰浓度之比和暴露量之比也与给药剂量呈正比,2B11注射液在体内基本呈线性动力学特征。结论在本实验条件下,恒河猴2周重复iv给予2B113次是安全的,400 mg·kg^(-1)未观察到临床不良反应。
基金the National Natural Science Foundation of China(No.22236002)National Key R&D Program(Nos.2023YFF1105003 and 2022YFA1207300).
文摘Human epidermal growth factor receptor 2(HER2)is an important biomarker for detection and treatment of breast cancer.In this study,we developed monoclonal antibodies against the extracellular domain(ECD)of HER2 and established a rapid and accurate lateral flow immunoassay(LFIA)for use in community medical institutions.The gene sequence of human HER2-ECD was obtained from the National Center for Biotechnology Information(NCBI)to construct the expression plasmid.HER2-ECD protein expressed in HEK293F cells was used to immunize BALB/c mice.The monoclonal antibodies were produced in mouse ascites and isolated by hybridoma cell screening.Antibodies were analyzed for purity by SDS-PAGE(sodium dodecyl sulphate-polyacrylamide gel-electrophoresis)and affinity was assessed by enzyme-linked immunosorbent assay(ELISA)while subtypes were detected using the commercial kits.The HER2-ECD test strip was prepared based on the sandwich method and evaluated using a portable detection instrument.The affinity of the paired antibodies,4D8 and 8D9,both reached 1×108 L/mol.Both antibodies specifically recognized the HER2-ECD protein in serum.The limit of detection(LOD)of the gold nanoparticle(AuNP)-based LFIA was 1.7 ng/mL with a detection range of 1.7-400 ng/mL,and the performance of the HER2-ECD strip correlated well with that of a Siemens chemiluminescent immunoassay(CLIA)kit.In conclusion,the paired antibodies were successfully prepared with high affinity and specificity.The AuNP-based LFIA of HER2-ECD provides a fast and accurate method to detect the concentration of HER2-ECD in serum samples for clinical use in community medical institutions,and could contribute to determining the progress of the disease or the effectiveness of treatment.
