Tkachenko Alexander, Xie Huan, Franzen Stefan, Feldheim Daniel L
Department of Chemistry, North Carolina State University, Raleigh, USA.
Methods Mol Biol. 2005;303:85-99. doi: 10.1385/1-59259-901-X:085.
In this chapter, we outline protocols for assembling and characterizing peptide-gold nanoparticle conjugates. We describe two strategies for attaching peptides to gold nanoparticles. One involves the covalent coupling of cysteine-terminated peptides directly to a particle surface via a sulfur-gold bond. Alternatively, peptides are coupled to bovine serum albumin (BSA) via a bifunctional molecular crosslinker. We also describe a number of characterization methods for determining the number of crosslinkers per BSA, peptides per BSA, and peptides adsorbed per particle. Finally, we show that the enormous visible light extinction properties of gold nano particles make them excellent imaging agents for tracking the trajectories of peptides inside living cells.
在本章中,我们概述了组装和表征肽 - 金纳米颗粒缀合物的方案。我们描述了两种将肽连接到金纳米颗粒上的策略。一种是通过硫 - 金键将半胱氨酸末端的肽直接共价偶联到颗粒表面。另一种方法是通过双功能分子交联剂将肽偶联到牛血清白蛋白(BSA)上。我们还描述了一些表征方法,用于确定每个BSA上的交联剂数量、每个BSA上的肽数量以及每个颗粒吸附的肽数量。最后,我们表明金纳米颗粒巨大的可见光消光特性使其成为追踪活细胞内肽轨迹的优秀成像剂。