第一型、第二型及第三型膠原蛋白製備及評估技術

Preparation and Evaluation Techniques for Type Ⅰ, Ⅱ and Ⅲ Collagens Using in Biomedical Field

林詠凱(Yung-Kai Lin) ； 劉登城(Deng-Cheng Liu)

《中華農學會報》 7卷2期 (2006/04) Pp. 133-147

https://doi.org/10.6730/JAAC.200604_7(2).0002

摘要

第一型、第二型與第三型膠原蛋白分別具有獨特之生理活性，且廣泛應用於生醫領域；由於醫藥上應用常需純度(purity)較高之膠原蛋白，因此需利用各種分離與純化技術自動物組織中製備各類型膠原蛋白。本文探討第一型、第二型與第三型膠原蛋白分離與純化技術並列舉各種評估技術供相關產業之參考。不同濃度之氯化鈉、硫酸銨及乙醇等沈澱劑存在下，第一型、第二型及第三型繆原蛋白之溶解度均不相同；第三型膠原蛋白難以藉這些沈澱劑分離，但可利用變性－復性的方式可有效分離第一型及第三型膠原蛋白。液相層析技術可提高膠原蛋白純度，但易受限於容量及成本；近年來發展之新穎濾紙基材管柱(filter paper-based chromatography)技術可大幅提昇膠原蛋白純化之容量及效率。經分離或純化之膠原蛋白可進一步藉由胺基酸組成、電泳、熱變性溫度、體外纖維重組、光譜分析技術及酵素安定性等技術進行評估其做為生醫材料之可行性。

關鍵字

膠原蛋白 ； 分離 ； 純化 ； 特性分析

並列摘要

Type Ⅰ, Ⅱand Ⅲ collagens possess specific biological activity and widely utilize a number of biomedical fields. Nevertheless, a higher purity of three distinct collagens must be required. Thus, several techniques for purification and separation of native type Ⅰ, Ⅱ and Ⅲ collagens were reviewed and discussed in the present article. Differential solubility of type Ⅰ, Ⅱ and Ⅲ collagens were made in the presence of different precipitants such as sodium chloride, ammonium sulfate and ethanol. Type Ⅲ collagen was difficult to separate by differential precipitation methods, which could be separated by differential denature-renature treatments. The purity of collagens could be elevated by the method of liquid chromatography. This, however, is limited by cost and capacity. Recently, a novel filter paper-based (FPB) DEAE-cellulose was developed to improve capacity and efficiency of traditional chromatographic systems. Several techniques such as amino acid composition, electrophoresis, thermal stability, spectroscopic properties, in vitro fibrillogenesis test and enzymatic sensitivity have been used to evaluate these three distinct collagens for the possibility of biomedical material.

並列關鍵字

collagen ； separation ； purification ； characterization