Hacettepe University, Department of Chemistry, Beytepe, Ankara, Turkey.
Colloids Surf B Biointerfaces. 2013 Dec 1;112:1-8. doi: 10.1016/j.colsurfb.2013.07.010. Epub 2013 Jul 18.
In this study, we aimed to develop an alternative matrix able to deplete the albumin (Alb) and immunoglobulin G (IgG) from blood plasma simultaneously to prepare plasma samples for large-scale applications of blood-related proteomics. As a first step, nano-protein A nanoparticles (nanoProA) were prepared and characterized. Subsequently, cibacron blue F3GA (CB) was immobilized onto the nanoProA's to enhance their specific affinity for Alb molecules. Finally, both nanoparticles, specifically, nanoProA and CB-nanoProA, were separately embedded into cryogel structures to combine advantages of the nanoparticles with those of the cryogels. The protein adsorption was optimized using aqueous Alb and IgG solutions separately. Subsequently, competitive protein adsorption was performed using a protein mixture prepared with Alb and IgG adhering to their plasma protein ratios. Because of the CB-immobilization, the Alb depletion performance of the cryogels increased whereas the IgG depleting performance decreased. Using the nanoProA, embedded cryogel removed 99.3% of the IgG, while using the CB-nanoProA embedded cryogel removed 97.5% of the Alb content. The simultaneous depletion performances of the cryogels for Alb and IgG were characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In this study, the monolithic cryogel-based adsorbents were classified as an alternative matrix to prepare plasma samples for proteomics applications at the preparative scale.
在这项研究中，我们旨在开发一种替代基质，能够同时从血浆中耗尽白蛋白（Alb）和免疫球蛋白 G（IgG），以准备用于血液相关蛋白质组学的大规模应用的血浆样品。作为第一步，制备并表征了纳米蛋白 A 纳米颗粒（nanoProA）。随后，将氰基三嗪三嗪（CB）固定在 nanoProA 上，以增强其对 Alb 分子的特异性亲和力。最后，分别将两种纳米颗粒（即 nanoProA 和 CB-nanoProA）嵌入到冷冻凝胶结构中，以结合纳米颗粒和冷冻凝胶的优点。使用单独的水性 Alb 和 IgG 溶液优化蛋白质吸附。随后，使用根据其在血浆蛋白质中的比例附着的 Alb 和 IgG 的蛋白质混合物进行竞争蛋白质吸附。由于 CB 固定化，冷冻凝胶的 Alb 耗尽性能提高，而 IgG 耗尽性能降低。使用 nanoProA 嵌入的冷冻凝胶可去除 99.3％的 IgG，而使用 CB-nanoProA 嵌入的冷冻凝胶可去除 97.5％的 Alb 含量。使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对冷冻凝胶的 Alb 和 IgG 的同时耗尽性能进行了表征。在这项研究中，基于整体的冷冻凝胶吸附剂被归类为用于在制备规模上进行蛋白质组学应用的血浆样品制备的替代基质。
