Sequence-specific retention calculator. A family of peptide retention time prediction algorithms in reversed-phase HPLC: Applicability to various chromatographic conditions and columns
Sequence-specific retention calculator. A family of peptide retention time prediction algorithms in reversed-phase HPLC: Applicability to various chromatographic conditions and columns
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DOI：
10.1021/ac071474k
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发表时间：
2007-11-15
影响因子：
7.4
通讯作者：
Krokhin, Oleg V.
中科院分区：
文献类型：
--
作者：
Spicer, Vic;Yamchuk, Andriy;Krokhin, Oleg V.
比较了不同制造商的C18反相色谱柱的分离选择性，以评估我们的序列特异性保留计算器（SSRCalc）肽保留预测算法的适用性。目前有三种不同版本的SSRCalc在用途：300埃孔径吸附剂（TFA作为离子配对改性剂，pH 2）、100埃（TFA，pH 2）和100埃（pH 10），它们已被应用于分离随机选择的胰蛋白酶肽混合物。影响C18吸附剂分离选择性的主要因素被认为是表观孔径，而封端化学的差异并没有引入显着的影响。将嵌入的极性基团引入C18官能团增加了含有具有极性基团Tyr和Trp的疏水氨基酸残基的肽的保留。我们还表明，改变离子配对改性剂甲酸/乙酸显着降低了算法的预测能力，因此，为不同的洗脱液条件开发的模型不能直接相互比较。
Separation selectivity of C18 reversed-phase columns from different manufacturers has been compared to evaluate the applicability of our sequence-specific retention calculator (SSRCalc) peptide retention prediction algorithms. Three different versions of SSRCalc are currently in use: 300-angstrom pore size sorbents (TFA as ion-pairing modifier, pH 2), 100 angstrom (TFA, pH 2), and 100 A (pH 10), which have been applied for the separation of randomly chosen mixture of tryptic peptides. The major factor affecting separation selectivity of C18 sorbents was found to be apparent pore size, while differences in end-capping chemistry do not introduce a significant impact. The introduction of embedded polar groups to the C18 functionality increases the retention of peptides containing hydrophobic amino acid residues with polar groups: Tyr and Trp. We also demonstrate that changing the ionpairing modifier to formic/acetic acid significantly reduces the algorithm's predictive ability, so models developed for different eluent conditions cannot be compared directly to each other.