目的:制备抗人前列腺特异性抗原(Prostate specific antigen,PSA)单克隆抗体。方法:利用盐析、离子交换和凝胶过滤的方法,从人精液中纯化前列腺特异性抗原,采用SDS-PAGE和游离前列腺特异性抗原定量试剂盒联合检测目的蛋白的纯度和含量,...目的:制备抗人前列腺特异性抗原(Prostate specific antigen,PSA)单克隆抗体。方法:利用盐析、离子交换和凝胶过滤的方法,从人精液中纯化前列腺特异性抗原,采用SDS-PAGE和游离前列腺特异性抗原定量试剂盒联合检测目的蛋白的纯度和含量,用纯化的PSA抗原免疫BALB/c小鼠,按常规的细胞融合与杂交瘤细胞的筛选方法,并对杂交瘤细胞产生的抗体进行了亚型、亲和力、特异性鉴定。结果:获得分泌抗-PSA单克隆抗体的杂交瘤细胞系15株,亲和力高,特异性和稳定性较好,亚型鉴定一株为IgG2a,其余均为IgG1。结论:本次制备的前列腺特异抗原单克隆抗体可用于临床PSA-ELISA诊断试剂的研发。展开更多
[Objective] TO study the combined extracting process of porcine blood superoxide dismutase （SOD） and other bioactive substances and thus to provide technical basis for making full use of blood resources and large-sc...[Objective] TO study the combined extracting process of porcine blood superoxide dismutase （SOD） and other bioactive substances and thus to provide technical basis for making full use of blood resources and large-scale production of SOD. [Method] Fibronectin, immunoglobulin, and hemoglobin were isolated from porcine blood, and SOD was extracted. Trace pyrogallol self-oxidation method to determine SOD activity was modified by optimizing the volume of pyrogallol and SOD samples, reaction temperature, and buffer pH. The specific activity of SOD was determined with the optimized extraction conditions. [ Result] The improved experimental conditions of SOD activity detection were as follows： 7 pyrogallol （50 mmol/L）, 3 ml Tris-HCI （50 mmol/L, pH 8.2）, reactive temperature at 25（3, and 10 pl SOD sample solution. The specific activity of extracted SOD was 5 056 U/mg protein. [ Conclusion] Four kinds of bioactive substance can be isolated from porcine blood by modern biological engi- neering integration technology, and the extracted SOD has better activity.展开更多
文摘目的:制备抗人前列腺特异性抗原(Prostate specific antigen,PSA)单克隆抗体。方法:利用盐析、离子交换和凝胶过滤的方法,从人精液中纯化前列腺特异性抗原,采用SDS-PAGE和游离前列腺特异性抗原定量试剂盒联合检测目的蛋白的纯度和含量,用纯化的PSA抗原免疫BALB/c小鼠,按常规的细胞融合与杂交瘤细胞的筛选方法,并对杂交瘤细胞产生的抗体进行了亚型、亲和力、特异性鉴定。结果:获得分泌抗-PSA单克隆抗体的杂交瘤细胞系15株,亲和力高,特异性和稳定性较好,亚型鉴定一株为IgG2a,其余均为IgG1。结论:本次制备的前列腺特异抗原单克隆抗体可用于临床PSA-ELISA诊断试剂的研发。
文摘[Objective] TO study the combined extracting process of porcine blood superoxide dismutase （SOD） and other bioactive substances and thus to provide technical basis for making full use of blood resources and large-scale production of SOD. [Method] Fibronectin, immunoglobulin, and hemoglobin were isolated from porcine blood, and SOD was extracted. Trace pyrogallol self-oxidation method to determine SOD activity was modified by optimizing the volume of pyrogallol and SOD samples, reaction temperature, and buffer pH. The specific activity of SOD was determined with the optimized extraction conditions. [ Result] The improved experimental conditions of SOD activity detection were as follows： 7 pyrogallol （50 mmol/L）, 3 ml Tris-HCI （50 mmol/L, pH 8.2）, reactive temperature at 25（3, and 10 pl SOD sample solution. The specific activity of extracted SOD was 5 056 U/mg protein. [ Conclusion] Four kinds of bioactive substance can be isolated from porcine blood by modern biological engi- neering integration technology, and the extracted SOD has better activity.
