Guo Xun, Chan-Park Mary B, Yoon Soon Fatt, Chun Jung-hoon, Hua Lin, Sze Newman S-K
Singapore-MIT Alliance, N2-B2c-15, Nanyang Technological University, 50 Nanyang Avenue, Singapore 639798.
Anal Chem. 2006 May 15;78(10):3249-56. doi: 10.1021/ac051773e.
This paper demonstrates a ultraviolet (UV)-embossed polymeric chip for protein separation by capillary isoelectric focusing (CIEF) and identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The polymeric chip was replicated by a UV-embossing technique using a soft rubber mold. Five diverse widely investigated families of UV-cured formulations were examined for MALDI ionization efficiency of bovine serum albumin (BSA) samples spotted on the polymer surfaces. The signal and signal-to-noise ratio from the polyester formulation were each 12 times those obtained with PMMA (a commonly used polymer material for MALDI) at picomole sample concentration. A polyester chip was successfully used to carry out CIEF to separate proteins, followed by MALDI-TOF-MS identification. Issues related to the successful chip fabrication and protein separation and identification are discussed.
本文展示了一种用于蛋白质分离和鉴定的紫外(UV)压纹聚合物芯片,该芯片通过毛细管等电聚焦(CIEF)进行蛋白质分离,并通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)进行鉴定。该聚合物芯片是使用软橡胶模具通过UV压纹技术复制而成的。研究了五种广泛研究的UV固化配方,用于检测点样在聚合物表面的牛血清白蛋白(BSA)样品的MALDI电离效率。在皮摩尔样品浓度下,聚酯配方的信号和信噪比分别是聚甲基丙烯酸甲酯(PMMA,一种常用于MALDI的聚合物材料)的12倍。成功使用聚酯芯片进行CIEF分离蛋白质,随后进行MALDI-TOF-MS鉴定。讨论了与芯片成功制造以及蛋白质分离和鉴定相关的问题。