Derivatized cellulose combined with MALDI-TOF MS: A new tool for serum protein profiling
Derivatized cellulose combined with MALDI-TOF MS: A new tool for serum protein profiling
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DOI：
10.1021/pr050227z
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发表时间：
2005-11-01
影响因子：
4.4
通讯作者：
Bonn, GK
中科院分区：
文献类型：
--
作者：
Feuerstein, I;Rainer, M;Bonn, GK
为从各种生物样本中发现生物标记物提供一种快速、灵敏的蛋白质谱方法对于引入新的标记物至关重要，这些标记物可以改善癌症患者在肿瘤早期的诊断，从而增加治疗的机会。我们在这里报告衍生化纤维素颗粒的开发和应用，用于选择性的血清蛋白质图谱。在固定化金属离子亲和层析(IMAC)中，纤维素先用甲基丙烯酸缩水甘油酯(GMA)衍生，然后用亚氨基二乙酸(IDA)衍生化。为了探索这种材料在人血清样品蛋白质谱中的应用，将血清样品与衍生的纤维素颗粒一起搅拌到悬浮液中，在30℃下孵育2小时，洗涤后，将1亩L的IDA-Cu2+-纤维素悬液直接涂在MALDI-靶标上，与芥子酸(SA)混合，并用MALDI-TOF MS分析，从多次分析的等度样品中获得一致的血清特异性数据，表明该方法的可靠性。然而，所获得的血清指纹被证明对任何给定的血清都是特异的。该技术允许样品在显影的靶上高度浓缩，从而获得高灵敏度和重现性，而不会耗尽白蛋白和免疫球蛋白，并在MS分析之前样品洗脱。这项研究首次证明，衍生纤维素颗粒与MALDI-TOF MS相结合，代表了一种简单、经济和快速的方法来生成用于生物标记物鉴定的血清蛋白质图谱。
Providing a rapid and sensitive protein profiling method for biomarker discovery from a variety of biological samples is crucial for the introduction of new markers that improve cancer patient diagnosis at early tumor stages, thus increasing the chances of curative treatment. We report here the development and application of derivatized cellulose particles for selective serum protein profiling. For immobilized metal ion affinity chromatography (IMAC), cellulose was derivatized with glycidyl methacrylate (GMA) and subsequently with iminodiacetic acid (IDA). To investigate the application of this material for generating protein profiles of human serum samples, the serum samples were agitated with the derivatized cellulose particles to a suspension and incubated for 2 h at 30 degrees C. After washing, 1 mu L of the IDA-Cu2+-cellulose suspension was applied directly onto a MALDI-target, mixed with sinapinic acid (SA) and analyzed with MALDI-TOF MS. Consistent serum specific data were obtained from aliquoted samples analyzed several times, indicating the reliability of the method. However, the serum fingerprints obtained proved to be specific for any given serum. The technique presented allows a high enrichment of sample on the developed target leading to a high sensitivity and reproducibility without depletion of albumin and immunoglobulin, and sample elution prior to MS-analysis. The study demonstrates for the first time that derivatized cellulose particles combined with MALDI-TOF MS represent a simple, economical, and rapid approach to generate serum protein profiles for biomarker identification.