• 作者： Chen, W.Y.; Liu, P.K.; Wang, L.K.et al.
• 中文摘要： 本研究的目的為根據已知胺基酸順序的胜或蛋白質,以分子辨識理論、蛋白質合成原理及Kyte與Doolittle的親疏水值系統,設計出 Antisense胜或蛋白質作為親和性吸附分離觸手。本研究選擇親水性的Angiotensin II(AII:Asp- Arg-Tyr-Ile-His-Pro-Phe)當成目標胜,根據本研究所發展的分子辨識理論觸手設計軟體所設計的反意胜觸手則為AII-AS(Ile-Ala-Asn-Val-Asn-Met-Gly- Glu)。此結果與由AII DNA鹽基順序所解譯的 Antisense胜有相似的胺基酸親疏水性分布。將所合成之AII-AS固定在EAH-Sepharose 4B親和性吸附劑且分別對AII及AII的阻抗劑(Antagonist)作批次的恆溫吸附實驗,並以Langmuir isotherm描述,得其與AII-AS之結合常數為728.99(L/mole AII)及 265.58(L/mole AII antagonist)。並且以Biosym公司之電腦分子模擬軟體Insight II,計算互相結合的胜可能產生的氫鍵數目。在定性上顯示AII antagonist對固定化之AII-AS的作用力比AII強。最後並以親和性管柱吸附曲線的數學模式來描述 CAS-AII層柱層析之行為,並探討操作參數的改變,對於此吸附反應管柱層析的影響,結果發現孔隙擴散在本親和性管柱吸附中為速率控制步驟。
• 英文摘要： An "antisense" peptide corresponding to the "sense" peptide, angiotensin II, was designed by the Molecular Recognition Theory (MRT) and synthesized by the solid phase peptide synthesis method. The complementarity of peptides specified by "sense" and "antisense" strands of DNA of MRT were adapted in a computer program and the Kyte and Doolittle's hydropathy scoring system was used as the hydrophobicity indicator of amino acids in this study. The prototypical system used for evaluating the interaction between such peptide pairs involved testing for the binding of angiotensin II (AII) and its antisense peptide, AII-AS(Ile-Ala-Asn-Val-Asn-Met-Gly-Glu). AII-AS peptide was attached to EAH-Sepharose 4B support, and the binding constants between AII-AS with AII and with AII antagonist( [ Ser/sup 1/, Ile/sup 8/ ] -AII) were respectively determined. The higher binding affinity between AII antagonist with AII-AS was also supported by a preliminary molecular modeling calculation. A mathematical modeling for affinity chromatography was adapted to compare the experimental results and to demonstrate the effects of the pore diffusion as the rate determining step on the performance of the anti-sense affinity column. The results of this study could provide a novel approach to affinity ligand design for bioseparation by MRT and molecular modeling.
• 中文關鍵字： 分子辨識理論; 胜; 配位基; 親和性層析; 純化
• 英文關鍵字： Molecular Recognition Theory; Peptide; Ligand; Affinity Chromatography; Purification