Scaffold Diversification Enhances Effectiveness of a Superlibrary of Hyperthermophilic Proteins
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- 作者：Mahmud Hussain ; Nimish Gera ; Andrew B. Hill ; Balaji M. Rao
- 刊名：ACS Synthetic Biology
- 出版年：2013
- 出版时间：January 18, 2013
- 年：2013
- 卷：2
- 期：1
- 页码：6-13
- 全文大小：497K
- 年卷期：v.2,no.1(January 18, 2013)
- ISSN：2161-5063
文摘
The use of binding proteins from non-immunoglobulin scaffolds has become increasingly common in biotechnology and medicine. Typically, binders are isolated from a combinatorial library generated by mutating a single scaffold protein. In contrast, here we generated a 鈥渟uperlibrary鈥?or 鈥渓ibrary-of-libraries鈥?of 4 脳 108 protein variants by mutagenesis of seven different hyperthermophilic proteins; six of the seven proteins have not been used as scaffolds prior to this study. Binding proteins for five different model targets were successfully isolated from this library. Binders obtained were derived from five out of the seven scaffolds. Strikingly, binders from this modestly sized superlibrary have affinities comparable or higher than those obtained from a library with 1000-fold higher sequence diversity but derived from a single stable scaffold. Thus scaffold diversification, i.e., randomization of multiple different scaffolds, is a powerful alternate strategy for combinatorial library construction.