Department of Chemistry, University of Western Ontario, London, ON, Canada.
J Mass Spectrom. 2010 Mar;45(3):241-51. doi: 10.1002/jms.1708.
A novel approach to high-throughput sequence deconvolution of on-bead small peptides (MW < 2000 Da) using on-target MALDI-TOF/TOF instrumentation is presented. Short peptides of pentamer and octamer length, covalently attached to TentaGel polystyrene beads through a photolabile linker, were placed onto the MALDI target, apportioned with suitable matrix (2,5-dihydroxybenzoic acid) and then hit with the instrument laser (Nd : YAG, 355 nm). This induced easy and highly reproducible photochemical cleavage, desorption (MS mode) and fragmentation (MS/MS mode). Peptide fragments were identified with a mass accuracy of 0.1 Da of the expected values. This technique significantly accelerates the sequence determination of positive peptide hits obtained from random combinatorial libraries when screening against biological targets, paving the way for a rapid and efficient method to identify molecular imaging ligands specific to pathological targets in cancer and other diseases.
本文提出了一种新的高通量方法，用于通过在靶 MALDI-TOF/TOF 仪器对在珠小肽（MW < 2000 Da）进行高通量序列反卷积。通过光不稳定连接子共价连接到 TentaGel 聚苯乙烯珠上的五聚体和八聚体长度的短肽被放置在 MALDI 靶上，用合适的基质（2,5-二羟基苯甲酸）分配，然后用仪器激光（Nd：YAG，355nm）照射。这诱导了简单且高度可重复的光化学裂解、解吸（MS 模式）和片段化（MS/MS 模式）。肽片段的鉴定具有预期值 0.1Da 的质量精度。当针对生物靶标筛选时，该技术可显著加速从随机组合文库中获得的阳性肽命中的序列确定，为识别癌症和其他疾病中病理靶标特异性分子成像配体的快速有效的方法铺平了道路。
