重组人酸性成纤维细胞生长因子C-末端测序

C-terminal sequencing of recombinant human acid fibroblast growth factor

目的:应用蛋白质 N-末端序列分析仪测定重组人酸性成纤维细胞生长因子(aFGF)C-末端氨基酸序列。方法:应用多肽分析软件选择合适的蛋白内切酶完全酶切 aFGF,酶切后用 RP-HPLC 进行分离,收集软件预测 C-末端肽段保留时间处的肽段峰,用蛋白质 N-末端序列分析仪直接测得该肽段氨基酸全序列。结果:实测肽图图谱与软件预测理论图谱一致,所得到的肽段为 aFGF C-末端肽段,经测序其结果与理论序列完全一致。结论:通过选择合适的蛋白内切酶,可运用 RP-HPLC和蛋白质 N-末端序列分析仪测定基因工程产品 C-末端氨基酸序列。

Objective：To sequence the C -terminal of recombinant human acid fibroblast growth factor （aFGF）. Method ：aFGF was completely digested by proper endoproteinase which was chosen by the software（ Peptide Companion）. The digested peptides were separated and purified through RP- HPLC. Then the peptide whose retention time was consistent with that of C -terminal peptide predicted by the software was collected and sequenced through PRO- TEIN SEQUENCER. Result：The peptide mapping was consistent with that predicted by the software. The obtained sequence was consistent with the theoretical C -terminal sequence of aFGF. Conclusion：The bio -pharmaceuticals C -terminal can be sequenced through proper endoproteinase digestion, RP -HPLC and PROTEIN SEQUENCER.