Department of Pharmaceutical Chemistry, School of Pharmacy, University of Oslo , NO-0316 Oslo, Norway.
Anal Chem. 2014 Dec 16;86(24):12291-8. doi: 10.1021/ac503559c. Epub 2014 Nov 26.
Biomarker mass spectrometry assays are in high demand, and analysis of pro-gastrin releasing peptide (ProGRP) as a small cell lung cancer marker has been recently investigated by mass spectrometry after immunoextraction. In this article, we introduce an assay based on molecularly imprinted polymers (MIPs) targeting the proteotypic peptide of ProGRP as a possible alternative to current immuno-based assay. The MIPs were prepared by surface-initiated reversible addition-fragmentation chain transfer polymerization and were introduced as sorbents for the cleanup and enrichment of a ProGRP signature peptide from tryptically treated serum samples. The use of an appropriate solid-phase extraction protocol allowed specific extraction of the target peptide while depleting other peptides that arose from the sample digestion, hence resulting in reduced background. The selective extraction of a ProGRP signature peptide, after digestion of serum samples, translates into a time- and cost-effective method suited for bottom-up analysis wherever targeted peptide extraction from complex matrices is required.
生物标志物质谱分析需求量很大，最近已经通过免疫提取后进行质谱分析来研究胃泌素释放肽前体（ProGRP）作为小细胞肺癌标志物。在本文中，我们介绍了一种基于针对 ProGRP 蛋白原肽的分子印迹聚合物（MIP）的测定法，作为当前基于免疫的测定法的可能替代方法。MIPs 是通过表面引发可逆加成-断裂链转移聚合制备的，并被用作从胰蛋白酶处理的血清样品中进行 ProGRP 特征肽的清洗和富集的吸附剂。使用适当的固相萃取方案可以特异性地提取目标肽，同时耗尽来自样品消化的其他肽，从而降低背景。在消化血清样品后，选择性地提取 ProGRP 特征肽可转化为一种节省时间和成本的方法，适用于需要从复杂基质中靶向提取肽的自下而上的分析。
