Bobály Balázs, Mikola Vivien, Sipkó Enikő, Márta Zoltán, Fekete Jenő
Department of Inorganic and Analytical Chemistry, Budapest University of Technology and Economics, Szt. Gellért tér 4, Budapest 1111, Hungary.
Department of Inorganic and Analytical Chemistry, Budapest University of Technology and Economics, Szt. Gellért tér 4, Budapest 1111, Hungary
J Chromatogr Sci. 2015 Aug;53(7):1078-83. doi: 10.1093/chromsci/bmu169. Epub 2014 Dec 11.
It was found that recoveries of proteins depend on trifluoroacetic acid concentration in the mobile phase and showed maximum in the range of 0.01-0.1 v/v%. Transferrin and lysozyme were used to evaluate the recoveries of proteins from dedicated reversed-phase columns. Different types of reversed-phase columns were evaluated, such as core shell type materials (Aeris Widepore with C4, C8 and C18 modification) as well as fully porous hybrid particles (Waters BEH, modified with C4 and C18 alkyl chains). Recoveries ranged between 60.7-95.2% for transferrin and 72.1-99.8% for lysozyme. Based on the data presented, at least two different adsorption effects, the well-known hydrophobic and silanophilic/polar interaction might influence the recovery. In addition to this, conformational effects due to ion pairing with the acidic mobile phase additive might change them.
研究发现，蛋白质的回收率取决于流动相中三氟乙酸的浓度，在0.01 - 0.1 v/v%范围内回收率最高。使用转铁蛋白和溶菌酶来评估专用反相柱中蛋白质的回收率。评估了不同类型的反相柱，如核壳型材料（Aeris Widepore，经C4、C8和C18修饰）以及全多孔杂化颗粒（Waters BEH，经C4和C18烷基链修饰）。转铁蛋白的回收率在60.7 - 95.2%之间，溶菌酶的回收率在72.1 - 99.8%之间。根据所呈现的数据，至少两种不同的吸附作用，即众所周知的疏水作用和亲硅烷化/极性相互作用可能会影响回收率。除此之外，与酸性流动相添加剂的离子配对引起的构象效应可能会改变回收率。
