【摘要】 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。更多还原

【Abstract】 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 Compan- ion).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.更多还原