以虾致敏蛋白Pen a 1（Tropomyosin）抗原表位为研究对象,建立了利用Pena 1表位抗体亲和纯化致敏蛋白的新方法.Fmoc法合成致敏Pen a 1蛋白的C端含有3个抗原表位的第247 ～284位氨基酸对应的多肽片段,应用马来酰亚胺法将多肽与KLH（匙孔...以虾致敏蛋白Pen a 1（Tropomyosin）抗原表位为研究对象,建立了利用Pena 1表位抗体亲和纯化致敏蛋白的新方法.Fmoc法合成致敏Pen a 1蛋白的C端含有3个抗原表位的第247 ～284位氨基酸对应的多肽片段,应用马来酰亚胺法将多肽与KLH（匙孔血蓝蛋白）、BSA（牛血清白蛋白）偶联制备人工免疫抗原（Pep-tide-KLH）和人工包被抗原（Peptide-BSA）,免疫人工抗原免疫纯种新西兰白兔,获得多克隆抗血清,抗血清经辛酸-硫酸铵及特异性血清纯化预装柱（HiTrap rProtein A FF）纯化后与溴化氰活化琼脂糖凝4B（CNBr-Activated Sepharose 4B）进行偶联.ELISA（酶联免疫吸附试验）测定该多克隆抗体效价为2.05×106,多肽对抗体的IC50（50％抑制浓度）为0.21 mg/L,交叉试验表明该抗体与虾中非Pena 1蛋白无交叉反应性;Bradford法测定CNBr-Activated Sepharose 4B与抗体的偶联率为90.76％.间接竞争ELISA测定1 mL偶联介质的吸附容量为2.84 mg Pen a 1,免疫亲和柱的加标回收率为89.6％～93.6％,亲和柱使用寿命为4次.展开更多
Dip-pen nanolithography(DPN) has been developed to pattern monolayer film of various molecules in submicrometer dimensions through the controlled movement of ink-coated atomic force microscopy(AFM) tip on a desired su...Dip-pen nanolithography(DPN) has been developed to pattern monolayer film of various molecules in submicrometer dimensions through the controlled movement of ink-coated atomic force microscopy(AFM) tip on a desired substrate, which makes DPN a potentially powerful tool for making the functional nanoscale devices. In this letter, using direct-write dip-pen nanolithography to generate nanoscale patterns of poly-L-lysine on mica was described. Poly-L-lysine molecules can anchor themselves to the mica surface through electrostatic interaction force, so stable poly-L-lysine patterns, such as square, line, circle and cross, could be obtained on freshly cleaved mica surface. From AFM image of the patterned poly-L-lysine nanostructures on mica, we know that poly-L-lysine was flatly bound to the mica surface. These oriented patterns of poly-L-lysine on mica can provide the prospect of building functional nanodevices and offer new options for this technique in a variety of other significant biomolecules.展开更多
文摘Dip-pen nanolithography(DPN) has been developed to pattern monolayer film of various molecules in submicrometer dimensions through the controlled movement of ink-coated atomic force microscopy(AFM) tip on a desired substrate, which makes DPN a potentially powerful tool for making the functional nanoscale devices. In this letter, using direct-write dip-pen nanolithography to generate nanoscale patterns of poly-L-lysine on mica was described. Poly-L-lysine molecules can anchor themselves to the mica surface through electrostatic interaction force, so stable poly-L-lysine patterns, such as square, line, circle and cross, could be obtained on freshly cleaved mica surface. From AFM image of the patterned poly-L-lysine nanostructures on mica, we know that poly-L-lysine was flatly bound to the mica surface. These oriented patterns of poly-L-lysine on mica can provide the prospect of building functional nanodevices and offer new options for this technique in a variety of other significant biomolecules.
