一、技術名稱：蛋白質純化的載體、方法、系統與試劑盒-美國專利

VECTORS, METHODS, SYSTEMS AND KITS FOR PROTEIN PURIFICATION

技術簡介

Core Techniques of patent US 8,236,529 B2

We developed a system for affinity purification with a cheap solid support and also possibly to eliminate the treatment of protease to retain the target protein. As chitin, a linear polymer of β-(1–4)-linked N-acetylglucosamine, is an abundant natural material produced in the biosphere and exhibits substantial mechanical and chemical strength, it is a perfect candidate to low the cost of protein purification. Three crystalline forms of chitin have been described in terms of the arrangement of the chitin chains: α-chitin in which chains run anti-parallel, β-chitin in which chains run parallel, and γ-chitin with mixed parallel/antiparallel chains. These natural polymers would provide an economical solid support for further protein purification if an appropriate binding apparatus were designed. In addition to a chitin-binding domain in chitinase, a chitin-binding protein (CBP) was found in the culture medium of Serratia marcescens when it was grown on chitin. This protein belongs to the carbohydrate-binding module (CBM) family 33 and is known to bind to β-chitin. Because the binding of CBP to chitin leads to non-hydrolytic disruption of the substrate structure, CBP can increase the substrate accessibility and strongly promote the hydrolysis of crystalline β-chitin by chitinase. Previous work showed also that CBP can bind to β-chitin at a pH about 8.0 and be eluted in acidic conditions, pH 7. This feature is prospectively useful for establishing a system for both protein purification and enzyme fixation. The construction of a fusion protein involves the linkage of two proteins or domains of proteins with a peptide linker. The selection of the linker sequence is particularly important for the construction of functional fusion proteins. Rigid α-helix peptide linkers (EAAAK)n (n≦6) are generally used for this purpose. These linkers are expected to form a monomeric hydrophilic α-helix, presumably stabilized by Glu––Lys+ salt bridges. We introduced a helix-forming peptide linker (EAAAK)5 between CBP and a target protein to discriminate effectively the bifunctional domains of a fusion protein.

 

美國專利 US 8,236,529 B2

專利權有效期間2009/12/18-2030/12/03

二、廠商資格：

1.與該技術領域相關之產業。

2.應有之研究或技術人員: 具相關領域之專業人員。

3.其他條件:無

4.應符合本校技術移轉相關規定。

 

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