重组人纽表位肽12的胃蛋白酶切肽图分析方法的研究

Peptide Mapping Analysis of Recombinant Human Neu Epitope Peptide 12 by Pepsin Digestion

采用反相高效液相色谱法建立重组人纽表位肽12的标准肽图分析法,固定相为十八烷基硅烷键合硅胶,流动相A液0.1%TFA的水溶液,流动相B液0.1%TFA的乙腈溶液;梯度洗脱程序0～70min内B%由0%增至70%。结果3批重组人纽表位肽12样品的反相高效液相色谱图完全一致,与重组人纽表位肽12理化对照品的肽图比较,完全吻合,同时胃蛋白酶并不干扰肽图的测定,该法准确度高、重现性好,可用于重组人纽表位肽12的质量控制。

To establish a standard method of peptide mapping analysis for quality control of recombinant human neu epitope peptide 12. Having found out the best pepsin digestion and chromatogram conditions by RP HPLC method. A C18 column was used and TFA acetonitrile water as mobile phase in the HPLC method. The detection wavelength was 214 nm . Peptide mappings of three continuous batches of recombinant human neu epitope peptide 12 and reference are completely identical. And pepsin does not interfere with peptide mapping. The method has high degree of accuracy and fine duplication which can be applied to quality control of recombinant human neu epitope peptide 12.