通过在合成DNA支架上组合配体来制造蛋白亲和试剂
Berea A R Williams1, Chris W Diehnelt, Paul Belcher
1Center for BioOptical Nanotechnology, Arizona State University, Tempe, Arizona 85287-5201, USA.
Journal of the American Chemical Society
|November 10, 2009
概括
研究人员开发了一种新的方法,通过结合小联体来制造合成抗体 (协同体). 这种方法避免了动物免疫,并为蛋白质组研究产生高亲和度蛋白质结合剂.
科学领域:
- 蛋白质组学是指蛋白质组学.
- 生物技术是生物技术.
- 合成生物学 合成生物学
背景情况:
- 了解蛋白质组需要所有蛋白质的亲和反应剂.
- 目前的抗体对大规模的蛋白质结合有局限性.
- 需要超越传统方法的新型蛋白亲和联结体.
研究的目的:
- 开发一种一般概念,用于制造无需动物免疫的蛋白亲和试剂.
- 通过在合成支架上组合配体,制造高亲和度双价试剂.
- 为了证明这种合成抗体方法的可行性.
主要方法:
- 选4000个独特的12-mer的图书馆与酵母蛋白Gal80.0结合.
- 利用DNA链接策略,在不同距离测试对.
- 通过组合最佳的联体,创建双价合成抗体 (协同体).
主要成果:
- 鉴定了可以结合到Gal80.0上非重叠位点的配体.
- 开发了一种对Gal80具有5nM亲和力的合成体,在ELISA和拉下测试中具有功能.
- 与单个相比,与双价同体结合的结合亲和力增加了1000倍.
结论:
- 合成抗体方法是产生高亲和度蛋白质结合剂的可行方法.
- 这种策略为全蛋白体应用提供了传统抗体生成的强大替代方案.
- 可以为各种目标创建同体,正如人类转基因所示.
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