老鼠免疫球蛋白IgG2a之位向性固定法—Fc區域的親和性配體設計;Peptide Ligand Design for Oriented Immobilization of Mouse IgG2a through Its Fc Region

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Title:	老鼠免疫球蛋白IgG2a之位向性固定法—Fc區域的親和性配體設計;Peptide Ligand Design for Oriented Immobilization of Mouse IgG2a through Its Fc Region
Authors:	曾偉志;Tseng,Wei-chih
Contributors:	化學工程與材料工程學系
Keywords:	表面電漿共振;分子動態模擬;分子嵌合;位向性固定;抗體;SPR;MD simulation;molecular docking;antibody;oriented immobilization
Date:	2013-08-28
Issue Date:	2014-07-24 13:51:22 (UTC+8)
Publisher:	國立中央大學
Abstract:	免疫生物感測器是藉由偵測在生物晶片表面上抗體(antibody)與抗原 (antigen)之間的辨識行為，以檢測溶液相中之有害物、病原體或是特定的標示物(marker)，而偵測的靈敏度會受到下列因素的影響：儀器感測訊號的機制、抗體在晶片表面的數量、抗體固定化的位向性。而抗體在晶片表面上隨機的固定化會造成偵測訊號下降，因此為了提升抗體抗原之間的辨識效率，抗體於基材表面的位向性固定化方式被廣為研究。我們提出一個策略設計與抗體 Fc 區域有高結合親和力(binding affinity)的胜肽配體(peptide ligand)。結合分子嵌合及分子動態模擬(molecular dynamics simulation，MD simulation)成功地設計、篩選出一條與Fc 區域有高親和力的胜肽配體，以此作為理想配體之主要序列並改質於金片表面上，使用表面電漿共振儀(surface plasmon resonance，SPR)量測Mouse IgG2a 在金片表面的吸附量，以及PSA 與Mouse IgG2a 的二抗(secondary antibody，2nd Ab)被辨識到的量，經計算後可得到Mouse IgG2a 的抗原辨識效率及位向因子。根據我們提出之設計策略，可設計出用於抗體位向性固定之胜肽配體，並改善抗體於表面之位向性，提升抗原辨識效率。 Immunosensors utilize specific bio-affinity interaction between antibody and antigen to detect toxicants, pathogens, or specific markers in complex mixtures. Antibodies need to be immobilized on the detection surface before targeted antigen can be captured. The recognition sensitivity depends on the sensitivity of signal sensing mechanism, the amount of immobilized antibody, and the orientation of antibody immobilized. Signal can be significantly reduced due to the random orientation of antibody on chip surface. Therefore, the methods for oriented immobilization of antibody were studied by many research groups in order to improve the recognition efficiency. We propose a strategy to design a small peptide ligand which has high binding affinity to the Fc region of antibody. It is expected that antibodies can be oriented immobilized on surface through the affinity between the desired Fc region and peptide ligand. Human prostate specific antigen (PSA) and PSA specific antibody derived from Mouse IgG2a are selected as the targeted antigen and antibody in this study. A peptide with a high Fc binding affinity is successfully designed and selected by combining the molecular docking and MD simulation. The recognition efficiency and orientation factor are measured by the adsorption amounts of PSA and the secondary antibody of Mouse IgG2a iii through the help of surface plasmon resonance (SPR). Our approach provided a new strategy for oriented immobilization of antibody by small ligands, which can significantly improve the recognition efficiency.
Appears in Collections:	[National Central University Department of Chemical & Materials Engineering] Electronic Thesis & Dissertation

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