Boireau W, Rouleau A, Lucchi G, Ducoroy P
Institut FEMTO-ST, Université de Franche Comté, Clinical-Innovation Proteomic Platform, 32 Av de I'Observatoire, CNRS, 25044 Besançon, France.
Biosens Bioelectron. 2009 Jan 1;24(5):1121-7. doi: 10.1016/j.bios.2008.06.030. Epub 2008 Jul 1.
We present the results of a study in which biomolecular interaction analysis (BIA, Biacoretrade mark 2000) was combined with mass spectrometry (MS) using entire "on-a-chip" procedure. Most BIA-MS studies included an elution step of the analyte prior MS analysis. Here, we report a low-cost approach combining Biacore analysis with homemade chips and MS in situ identification onto the chips without elution step. First experiments have been made with rat serum albumin to determine the sensitivity and validation of the concept has been obtained with an antibody/antigen couple. Our "on-a-chip" procedure allowed complete analysis by MS/MS(2) of the biochip leading to protein identifications at low femtomole to sub-femtomole levels. Using this technique, identification of protein complexes were routinely obtained giving the opportunity to the "on-a-chip" processing to complete the BIA-MS approach in the discovery and analysis of protein complexes.
我们展示了一项研究的结果，该研究采用完整的“芯片上”程序，将生物分子相互作用分析（BIA，Biacore商标2000）与质谱（MS）相结合。大多数BIA-MS研究在MS分析之前包括分析物的洗脱步骤。在此，我们报告了一种低成本方法，将Biacore分析与自制芯片相结合，并在芯片上进行原位质谱鉴定，无需洗脱步骤。首先用大鼠血清白蛋白进行了实验，以确定灵敏度，并用抗体/抗原对获得了该概念的验证。我们的“芯片上”程序允许通过MS/MS（2）对生物芯片进行完整分析，从而在低飞摩尔至亚飞摩尔水平上鉴定蛋白质。使用该技术，常规获得蛋白质复合物的鉴定结果，使“芯片上”处理有机会在蛋白质复合物的发现和分析中完善BIA-MS方法。
