本研究利用表面增顯紅外光譜儀(Surface-Enhanced Infrared Spectroscopy, SEIRAS)結合循環伏安儀(Cyclic Voltammetry, CV)，即時觀測牛血清白蛋白(Bovine serum albumin, BSA)在金電極表面的吸附行為，藉以探討六碳羧基(6-Mercaptohexanoic acid , 6-MHA)、六碳烷基(1-Hexanethiol, MCH)、六碳羥基(6-Mercapto-1-hexanol, 6-MCH)與兩碳氨基(Cysteamine , MEA)，不同官能基硫醇修飾，與不同電位之金表面，對BSA的吸附影響，再以FTIR分析吸附於金表面之BSA的構型變化，推測其吸附情況。實驗結果顯示，其總吸附量排序為：6-MHA > MCH > 6-MCH > MEA，推測為帶負電的羧基基團會吸引BSA分子的吸附。接著利用循環伏安儀使金表面帶有不同的電位，結果在施加負電位時的總吸附量會高於施加正電位的情況。以FTIR分析BSA之二級結構，在溶液中的構型以α-helix為主，而吸附於金表面時會轉變為β-sheet為主，但當BSA吸附量增加形成多層吸附層時，分子的構型以α-helix為主。
在實驗的另一部分使用6-MHA修飾金電極後，以抗體(Anti-HDGF)與抗原(HDGF)組裝成無標記型免疫生醫感測器，在抗原濃度為0.5~4 ng/c.c時，其誤差值約為 2.5~4.4% (n=5)。將檢測下限降低到0.125 ng/c.c時，檢量線以0.5 ng/c.c為轉折點，在0.125~0.5 ng/c.c的區間，電流密度與抗原濃度成正比，而在0.5~4 ng/c.c的區間則成反比。以FTIR分析抗原之構型，發現隨著抗原濃度的增加，β-sheet與α-helix之比例逐漸相近，當抗原濃度高於轉折點0.5 ng/c.c時，兩者之比例則越差越大。

In this study, surface-enhanced infrared spectroscopy (SEIRAS) combined with cyclic voltammetry (CV), was utilized to in-situ observe the adsorption behavior of Bovine serum albumin (BSA) on 6-Mercaptohexanoic acid (6-MHA), 1-Hexanethiol (MCH), 6-Mercapto-1-hexanol (6-MCH) and Cysteamine (MEA), different functional groups of the thiol modified gold, and different applying potential on gold surface. Using FTIR to analysis the conformation change of BSA which adsorbed to pure gold surface, and suggest it adsorption condition. The experimental results show that when different functional group modified gold surface, the order of total amount of adsorption is 6-MHA > MCH > 6-MCH > MEA, presume that negatively functional group would increase the amount of adsorption. Following, using CV to applying different potential on gold surface, the total amount of adsorption of applying negative potential is higher than applying positive potential. Using FTIR to analysis the conformation of BSA, in solution results is α-helix primarily, and it would convert to β-sheet primarily when adsorbed to gold surface, when the BSA gradually formed multilayer on the gold surface, the molecular is α-helix primarily.
At another part of experiment, we use 6-MHA to modified gold electrode, and use antibody (Anti-HDGF) and antigen (HDGF), assembled into label-free immunosensor, the deviation value is about  2.5~4.8 % (n=5) within the antigen range of 0.5~4 ng/c.c. As increasing the detect limit to 0.125 ng/c.c, the calibration curve with turning point is 0.5 ng/c.c, at 0.125~0.5ng/c.c region, the current density and antigen concentration are in direct proportion, and at 0.5~4 ng/c.c region, the current density and antigen concentration are in inverse proportion. Using FTIR to analysis the antigen conformation, as the antigen concentration increase, the ratio of β-sheet and α-helix is gradually close, when the antigen concentration is higher than the turning point of 0.5 ng/c.c, the difference of the ratio is more large.

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