Immobilization of enzymes and antibodies on magnetic particles isolated from magnetotactic bacteria

将酶和抗体固定在从趋磁细菌中分离的磁性颗粒上

• 批准号: 61470117
• 负责人: MATSUNAGA Tabashi
• 金额: $ 4.29万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61470117/
• 关键词: Magnetotactic bacteria; Magnetic particles; Immobilization; Glucose oxidase; Antigen-antibody reaction; 有機薄膜; マグネタイト; 固定化酵素

We have developed the novel use of magnetic particles isolated from magnetotactic bacteria. Magnetotactic bacteria were incubated at 25゜C for a fwe weeks in the sludge enriched with 0.005% sodium succinate and 0.005% sodium nitrate, and collected by use of a magnetic harvesting apparatus. Magnetotactic bacteria were mainly cocci and rod type. These bacteria contained about 10 to 20 magnetic particles. Biogenic magnetic particles were separated form magnetotactic bacteria by digestion with NaOH. The magnetic particles were 500-1000<ang> in size and aminly consist of magnetite (Fe_3O_4), which analysed by an energy dispersive X-ray analyzer and a highresolution transmission electron microscope. This particles were coverd with organic thin films.Immobilization of enzymes and antibodies on biogenic magnetic particles were carried out by treating magnetic particles with <gamma>-(aminopropyl)triethoxysilane and glutaraldehyde, and incubated with antibodies and enzymes at 4゜C for 12hr. Large … More amounts of enzymes and antibodies were immobilized on biogenic magnetic particles because of its uniform sharp and small size. Enzymes immobilized on magnetic particles show high activity.We measured glucose concentration by using glucose oxidase immobilized on magnetic particles and optic fiber sensor. Glucose concentration could be measured linearly in the range of 0.1-4.0 mg/ml. Enzyme reaction was enhanced by magnetic stirring. Selective separation of E.coli was also done by use of anti-E.coli antibody immobilized on magnetic particles. After magnetically stirring and separation, E.coli cell concentration decreased. Whereas the cell concentrations of L.acidphillus and S.cervisiae didn't decreased after magnetic separation. Therefore, particular cell could be separated selectivity. Furthermore, the small amount of antigen could be detected rapidly by microspopic observation of antigen-antibody reaction forming agglutination by use of the antibodies immobilized on magnetic particles. Biogenic magnetic particles are promising for the future application in clinical field. Less

我们已经开发了从趋磁细菌中分离的磁性颗粒的新用途。将趋磁细菌在富含0.005%琥珀酸钠和0.005%硝酸钠的污泥中于25 ° C培养四周，并使用磁性收集装置收集。趋磁细菌主要为球菌和杆状细菌。这些细菌含有大约10到20个磁性颗粒。用NaOH消化法从趋磁细菌中分离出生物磁性颗粒。用<ang>X射线能谱仪和高分辨透射电镜分析了磁性颗粒的组成，磁性颗粒的大小为500-1000 μ m，主要由磁铁矿（Fe_3O_4）组成。用氨基丙基三乙氧基硅烷和戊二醛处理生物磁性颗粒，将酶和抗体固定在生物磁性颗粒上<gamma>，并与抗体和酶在4 ℃孵育12小时。大 ...更多信息 由于生物磁性颗粒的均匀尖锐和小尺寸，大量的酶和抗体被固定在生物磁性颗粒上。利用磁性颗粒固定化葡萄糖氧化酶和光纤传感器对葡萄糖浓度进行了测量。葡萄糖浓度在0.1-4.0 mg/ml范围内呈线性。磁力搅拌对酶促反应有促进作用。还通过使用固定在磁性颗粒上的抗大肠杆菌抗体进行大肠杆菌的选择性分离。经磁力搅拌分离后，大肠杆菌细胞浓度下降。而嗜酸乳杆菌和啤酒酵母菌经磁分离后细胞浓度没有降低。因此，可以选择性地分离特定的细胞。此外，通过显微镜下观察抗原-抗体反应形成凝集反应，利用固定在磁性颗粒上的抗体可快速检测少量抗原。生物源性磁性微粒在临床上有很好的应用前景。少

项目成果

Tadashi Matsunaga: Biomagnetism.

松永正：生物磁学。

Minoru Funaki: J. Geomagnet. Geolectric.

Minoru Funaki：J. Geomagnet。

Tadashi,Matsunaga: "Isolation of Lymphocytes from Phagocytic Granulocyte and Macrophage Using Magnetotactic Bacteria and Magnetic Separation Technique" Biochim. Biophys. Acta.

Tadashi，Matsunaga：“使用趋磁细菌和磁分离技术从吞噬粒细胞和巨噬细胞中分离淋巴细胞”Biochim。

Minoru Funaki: "Identification of the Magnetic S Pole on the Selected Magnetic Grains from St. Severin Meteorite Using Magnetotactic Bacteria" J. Geomagnet. Geoelectric.

Minoru Funaki：“使用趋磁细菌识别圣塞维林陨石中选定磁性颗粒的磁 S 极”J. Geomagnet。

Tadashi Matsunaga: Appl. Microbiol. Biotechnol.26. 328-332 (1987)

松永正：应用。