Background A monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not c...Background A monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj).Methods A phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs select ed were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients.Results Six positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1.07×10 6 individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically an d had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60% and 37% in acute and chronic patients, respectively, with a specificity of 90%. When the two specific scFvs were combined, their sensitivity was found to be 75% and 57% in acute and chronic patients, respectively, with a specificity of 85%.Conclusions The results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construct ion also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.展开更多
经验取样法是一种有效的密集纵向研究数据收集方法。该方法的优点正逐渐被广大研究者所认同,国外运用经验取样法取得的研究成果越来越多,而国内学者对经验取样法的研究尚处于探索阶段,运用此方法的实证研究更是凤毛麟角。通过对《应用...经验取样法是一种有效的密集纵向研究数据收集方法。该方法的优点正逐渐被广大研究者所认同,国外运用经验取样法取得的研究成果越来越多,而国内学者对经验取样法的研究尚处于探索阶段,运用此方法的实证研究更是凤毛麟角。通过对《应用心理学期刊》（Journal of Applied Psychology）最近5年（2010-2014年）发表的26篇采用经验取样法的实证研究论文的内容分析,发现这些文章全面体现了经验取样法的特点、研究内容、样本量、激励方式、数据收集和操作过程等该方法的操作要素。对其进行总结不仅有利于对经验取样法在组织行为学中的应用状况进行总结,而且也能促进这一方法在未来的组织行为学和人力资源管理中的发展和应用。展开更多
基金ThisworkwassupportedbytheWHOSpecialProgramforResearchandTraininginTropicalDiseases (No 970 978) ,theScienceFoundationfromtheMinistryofHealthofChina (No 98 1 85 ),andtheGuangdongProvincialScienceFoundationfor"2 11"Engineering
文摘Background A monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj).Methods A phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs select ed were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients.Results Six positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1.07×10 6 individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically an d had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60% and 37% in acute and chronic patients, respectively, with a specificity of 90%. When the two specific scFvs were combined, their sensitivity was found to be 75% and 57% in acute and chronic patients, respectively, with a specificity of 85%.Conclusions The results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construct ion also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.
文摘经验取样法是一种有效的密集纵向研究数据收集方法。该方法的优点正逐渐被广大研究者所认同,国外运用经验取样法取得的研究成果越来越多,而国内学者对经验取样法的研究尚处于探索阶段,运用此方法的实证研究更是凤毛麟角。通过对《应用心理学期刊》（Journal of Applied Psychology）最近5年（2010-2014年）发表的26篇采用经验取样法的实证研究论文的内容分析,发现这些文章全面体现了经验取样法的特点、研究内容、样本量、激励方式、数据收集和操作过程等该方法的操作要素。对其进行总结不仅有利于对经验取样法在组织行为学中的应用状况进行总结,而且也能促进这一方法在未来的组织行为学和人力资源管理中的发展和应用。
