A Study on Fluorescence Spectroscopy of Immobilizing FITC Labeled Antibody between Fiber Optic Surface and Biomolecule
Author(s): CHAO Ke-fu, ZHANG You-lin, KONG Xiang-gui, ZENG Qing-hui, SONG Kai
Pages: 940-944
Year: 2007 Issue: 6
Journal: CHINESE JOURNAL OF LUMINESCENCE
Keyword: 光纤生物传感器; 物理吸附与共价键固定; 荧光光谱;
Abstract: 采用化学方法把FITC标记羊抗人抗体IgG共价固定到三氨基三乙氧基硅烷和戊二醛(APTES-Glu)修饰的石英光纤纤芯表面.通过研究未经过任何修饰的纤芯表面的吸附以及共价固定的FITC标记羊抗人抗体的荧光光谱性质发现:共价结合到纤芯表面的FITC荧光光谱相对于溶液中FITC的荧光峰位红移约9nm,而物理吸附的FITC的荧光峰位移动约4nm,而且两者相对荧光强度相差6倍.而在固定的人血清蛋白进行免疫反应后,FITC的荧光峰位只移动3～4nm,除此之外,研究了抗体共价键固定的稳定性问题.结果表明:共价键结合抗体的数量要大于表面吸附,共价键固定的FITC标记羊抗人抗体与光纤表面间的相互作用较吸附于光纤表面的FITC标记羊抗人抗体间的相互作用更强.这对于建立一种通过荧光光谱识别固体表面与生物分子的吸附还是共价结合的判据提供了物理基础.
Pages: 940-944
Year: 2007 Issue: 6
Journal: CHINESE JOURNAL OF LUMINESCENCE
Keyword: 光纤生物传感器; 物理吸附与共价键固定; 荧光光谱;
Abstract: 采用化学方法把FITC标记羊抗人抗体IgG共价固定到三氨基三乙氧基硅烷和戊二醛(APTES-Glu)修饰的石英光纤纤芯表面.通过研究未经过任何修饰的纤芯表面的吸附以及共价固定的FITC标记羊抗人抗体的荧光光谱性质发现:共价结合到纤芯表面的FITC荧光光谱相对于溶液中FITC的荧光峰位红移约9nm,而物理吸附的FITC的荧光峰位移动约4nm,而且两者相对荧光强度相差6倍.而在固定的人血清蛋白进行免疫反应后,FITC的荧光峰位只移动3～4nm,除此之外,研究了抗体共价键固定的稳定性问题.结果表明:共价键结合抗体的数量要大于表面吸附,共价键固定的FITC标记羊抗人抗体与光纤表面间的相互作用较吸附于光纤表面的FITC标记羊抗人抗体间的相互作用更强.这对于建立一种通过荧光光谱识别固体表面与生物分子的吸附还是共价结合的判据提供了物理基础.
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