AIM: To clone and express the antigen of monoclonal antibody (Mab) PD4 for further investigation of its function.METHODS: MGC803 cDNA expression library was constructed and screened with PD4 as probes to clone the ant...AIM: To clone and express the antigen of monoclonal antibody (Mab) PD4 for further investigation of its function.METHODS: MGC803 cDNA expression library was constructed and screened with PD4 as probes to clone the antigen. After failed in the library screening, immunoprecipitation and SDSpolyacrylamide gel electrophoresis were applied to purify the antigen for sequence analysis. The antigen coming from Nycoplasma hyorhinis (M. Hyorhinis) was further confirmed with Western blot analysis by infecting M. Hyorhinis-free HeLa cells and eliminating the M. Hyorhinis from MGC803cells. The full p37 gene was cloned by PCR and expressed successfully in Escherichia coli after site-directed mutations.Tmmunofluorescence assay was used to demonstrate if p37protein could directly bind to gastric tumor cell AGS.RESULTS: The cDNA library constructed with MGC803 cells was screened by Mab PD4 as probes. Unfortunately, the positive clones identified with Mab PD4 were also reacted with unrelated antibodies. Then, immunoprecipitation was performed and the purified antigen was identified to be a membrane protein of Mycoplasrna hyorhinis (M. Hyorhinis)by sequencing of N-terminal amino acid residues. The membrane protein was intensively verified with Western blot by eliminating M. Hyorhinis from MGC803 cells and by infecting M. Hyorhinis-free HeLa cells. The full p37 gene was cloned and expressed successfully in Escherichia coli after site-directed mutations. Immunofluorescence demonstrated that p37protein could directly bind to gastric tumor cell AGS.CONCLUSION: The antigen recognized by Mab PD4 is from M. Hyorhinis, which suggests the actions involved in Mab PD4 is possibly mediated by p37 protein or M. Hyorhinis. As p37 protein can bind directly to tumor cells, the pathogenic role of p37 involved in tumorigenesis justifies further investigation.展开更多
通过对Ag-4Pd键合合金线冷加工和热处理过程中的微观组织与性能进行研究,分析了冷变形加工率和热处理温度对Ag-4Pd键合合金线力学性能、组织结构和熔断电流的影响。研究结果表明:Ag-4Pd键合合金线随冷加工率增长,强度增加,伸长率降低,...通过对Ag-4Pd键合合金线冷加工和热处理过程中的微观组织与性能进行研究,分析了冷变形加工率和热处理温度对Ag-4Pd键合合金线力学性能、组织结构和熔断电流的影响。研究结果表明:Ag-4Pd键合合金线随冷加工率增长,强度增加,伸长率降低,滑移和孪生变形为主要形变类型;随着热处理温度的增加,?0.050 mm Ag-4Pd键合合金线拉断力下降,伸长率增加,525℃热处理时,Ag-4Pd键合合金线具有优秀的力学性能;进一步增加热处理温度,Ag-4Pd键合合金线出现孪晶组织,且孪生形核及亚晶吞并长大形核为主要形核方式;热处理过程中施加在线材上的拉紧力过大,导致Ag-4Pd键合合金线表面呈凹凸不平的微小竹节状;经试验数据拟合,Ag-4Pd键合合金线电阻值随热处理温度升高而降低,其熔断电流与熔断时间之间存在指数函数关系,Ag-4Pd键合合金线熔断电流与弧长之间存多项式函数关系。展开更多
基金Key Project of National Natural Science Foundation of China,No.30130190Beijing Natural Science Foundation,No.7012007Oncology Key Program and Cancer Center of Peking University
文摘AIM: To clone and express the antigen of monoclonal antibody (Mab) PD4 for further investigation of its function.METHODS: MGC803 cDNA expression library was constructed and screened with PD4 as probes to clone the antigen. After failed in the library screening, immunoprecipitation and SDSpolyacrylamide gel electrophoresis were applied to purify the antigen for sequence analysis. The antigen coming from Nycoplasma hyorhinis (M. Hyorhinis) was further confirmed with Western blot analysis by infecting M. Hyorhinis-free HeLa cells and eliminating the M. Hyorhinis from MGC803cells. The full p37 gene was cloned by PCR and expressed successfully in Escherichia coli after site-directed mutations.Tmmunofluorescence assay was used to demonstrate if p37protein could directly bind to gastric tumor cell AGS.RESULTS: The cDNA library constructed with MGC803 cells was screened by Mab PD4 as probes. Unfortunately, the positive clones identified with Mab PD4 were also reacted with unrelated antibodies. Then, immunoprecipitation was performed and the purified antigen was identified to be a membrane protein of Mycoplasrna hyorhinis (M. Hyorhinis)by sequencing of N-terminal amino acid residues. The membrane protein was intensively verified with Western blot by eliminating M. Hyorhinis from MGC803 cells and by infecting M. Hyorhinis-free HeLa cells. The full p37 gene was cloned and expressed successfully in Escherichia coli after site-directed mutations. Immunofluorescence demonstrated that p37protein could directly bind to gastric tumor cell AGS.CONCLUSION: The antigen recognized by Mab PD4 is from M. Hyorhinis, which suggests the actions involved in Mab PD4 is possibly mediated by p37 protein or M. Hyorhinis. As p37 protein can bind directly to tumor cells, the pathogenic role of p37 involved in tumorigenesis justifies further investigation.
文摘通过对Ag-4Pd键合合金线冷加工和热处理过程中的微观组织与性能进行研究,分析了冷变形加工率和热处理温度对Ag-4Pd键合合金线力学性能、组织结构和熔断电流的影响。研究结果表明:Ag-4Pd键合合金线随冷加工率增长,强度增加,伸长率降低,滑移和孪生变形为主要形变类型;随着热处理温度的增加,?0.050 mm Ag-4Pd键合合金线拉断力下降,伸长率增加,525℃热处理时,Ag-4Pd键合合金线具有优秀的力学性能;进一步增加热处理温度,Ag-4Pd键合合金线出现孪晶组织,且孪生形核及亚晶吞并长大形核为主要形核方式;热处理过程中施加在线材上的拉紧力过大,导致Ag-4Pd键合合金线表面呈凹凸不平的微小竹节状;经试验数据拟合,Ag-4Pd键合合金线电阻值随热处理温度升高而降低,其熔断电流与熔断时间之间存在指数函数关系,Ag-4Pd键合合金线熔断电流与弧长之间存多项式函数关系。
文摘利用扫描电镜、强度测试仪、热导率测试仪分析了Ag-4Pd和Ag-4Pd-0.5Ru键合合金线的性能及组织差异,研究了微量Ru元素对Ag-4Pd键合合金线球焊点尺寸、焊点形貌及键合强度的影响。结果表明:Ru元素的加入使Ag-4Pd键合合金线热导率由403 W/m?K降低到385 W/m?K,Ag-4Pd-0.5Ru键合合金线无空气焊球(free air ball)形状较Ag-4Pd键合合金线规则,提高了键合合金线焊点连接强度;Ag-4Pd键合合金线中的Ru元素使其热影响区长度由50μm减小至35μm,消除了由于热影响区长度过大导致的颈部微裂纹缺陷;Ag-4Pd键合合金线中的Ru元素细化了晶粒,增加了相同体积内不同取向的金属晶粒数目,改善了变形时的协同作用,降低了形变的不均匀性程度,获得了较规则的Ag-4Pd键合合金线球焊点形貌。
