The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography (HPAC) and quartz crystal microbalance (QCM) biosensor. Fragment 1-14 of human interferon-β (hIFN-...The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography (HPAC) and quartz crystal microbalance (QCM) biosensor. Fragment 1-14 of human interferon-β (hIFN-β) was chosen as sense peptide and its three antisense peptides (AS-IFN 1, AS-IFN 2, and AS-IFN 3) were designed according to the degeneracy of genetic codes. The affinity column was prepared with sense peptide as ligand and the affinity chromatographic behavior was evaluated. Glu-substituted antisense peptide (AS-IFN 3) showed the strongest binding to immobilized sense peptide at pH 7.5. A quartz crystal microbalance-flow injection analysis (QCM-FIA) system was introduced to investigate the recognition process in real-time. The equilibrium dissociation constants between sense peptide and AS-IFN 1, AS-IFN 2 and AS-IFN 3 measured 2.08×10-4, 1.31×10-4 and 2.22× 10-5 mol/L, respectively. The mechanism study indicated that the specific recognition between sense peptide and AS-IFN 3 was due to sequence-dependent and multi-modal affinity interaction.展开更多
A sensitive CD44-biosensor based on quartz crystal microbalance(QCM) was proposed for evaluating metastatic potential of breast cancer cells by using hyaluronan(HA) functionalized substrate film,polydopamine and polye...A sensitive CD44-biosensor based on quartz crystal microbalance(QCM) was proposed for evaluating metastatic potential of breast cancer cells by using hyaluronan(HA) functionalized substrate film,polydopamine and polyethyleneimine composite film,for the purpose of capturing CD44-positive cancer cells through specific binding of HA to CD44.Two differently CD44-expressed breast cancer cell lines(MDAMB-231 cells and MCF-7 cells) were put to use as targets for quantitative analysis as well as evaluation of metastatic potential of the cells.The limit of detection for MDA-MB-231(M231) cells and MCF-7 cells were 300 and 1,000 cells mL ~1 respectively.The expression level of CD44 on M231 cells exhibited two times higher than that of MCF-7 cells,indicating of a higher metastatic potential.Moreover,poly-Llysine modified QCM sensor was applied to monitor the stiffness of breast cancer cells that can reflect metastatic potential of cells.The results revealed that the MCF-7 cells were stiffer than M231 cells,implying that the M231 cells possessed higher metastatic potential.The proposed protocol is simple and rapid to evaluate the metastatic potential of cancer cells,in addition to offering a promising diagnostic tool for metastatic cancer.展开更多
基金Supported by the National Natural Science Foundation of China (Grant Nos. 20575072, 20435030 and 90408018)the Chinese Academy of Sciences (Grant No. KJCX2-SW-H06)
文摘The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography (HPAC) and quartz crystal microbalance (QCM) biosensor. Fragment 1-14 of human interferon-β (hIFN-β) was chosen as sense peptide and its three antisense peptides (AS-IFN 1, AS-IFN 2, and AS-IFN 3) were designed according to the degeneracy of genetic codes. The affinity column was prepared with sense peptide as ligand and the affinity chromatographic behavior was evaluated. Glu-substituted antisense peptide (AS-IFN 3) showed the strongest binding to immobilized sense peptide at pH 7.5. A quartz crystal microbalance-flow injection analysis (QCM-FIA) system was introduced to investigate the recognition process in real-time. The equilibrium dissociation constants between sense peptide and AS-IFN 1, AS-IFN 2 and AS-IFN 3 measured 2.08×10-4, 1.31×10-4 and 2.22× 10-5 mol/L, respectively. The mechanism study indicated that the specific recognition between sense peptide and AS-IFN 3 was due to sequence-dependent and multi-modal affinity interaction.
基金supported by the National Natural Science Foundation of China(21375048)
文摘A sensitive CD44-biosensor based on quartz crystal microbalance(QCM) was proposed for evaluating metastatic potential of breast cancer cells by using hyaluronan(HA) functionalized substrate film,polydopamine and polyethyleneimine composite film,for the purpose of capturing CD44-positive cancer cells through specific binding of HA to CD44.Two differently CD44-expressed breast cancer cell lines(MDAMB-231 cells and MCF-7 cells) were put to use as targets for quantitative analysis as well as evaluation of metastatic potential of the cells.The limit of detection for MDA-MB-231(M231) cells and MCF-7 cells were 300 and 1,000 cells mL ~1 respectively.The expression level of CD44 on M231 cells exhibited two times higher than that of MCF-7 cells,indicating of a higher metastatic potential.Moreover,poly-Llysine modified QCM sensor was applied to monitor the stiffness of breast cancer cells that can reflect metastatic potential of cells.The results revealed that the MCF-7 cells were stiffer than M231 cells,implying that the M231 cells possessed higher metastatic potential.The proposed protocol is simple and rapid to evaluate the metastatic potential of cancer cells,in addition to offering a promising diagnostic tool for metastatic cancer.
