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目的建立应用N-末端氨基酸测序数据估计N-末端不均一的单链重组蛋白制品主肽链比例的方法。方法利用氨基酸测序仪的摩尔归一化数据,经信号和噪音分离、信号的筛选等数据处理过程,将主肽链N-末端序列数据与其他次要序列数据分开,进而估计主肽链所占比例。将本方法计算结果与质谱结果进行比较,另用混合样品数据进行验证分析。结果本方法所计算结果基本符合实际情况,可较准确地估计主肽链比例。结论初步建立了应用N-末端氨基酸测序数据估计N-末端不均一的单链重组蛋白制品主肽链比例的方法,为相关蛋白的分析和研究提供了辅助。
OBJECTIVE: To establish a method for estimating the ratio of the major peptide chains of N-terminal non-uniform single-stranded recombinant protein products using N-terminal amino acid sequencing data. Methods Using the data normalized by amino acid sequencer, the N-terminal sequence of the main peptide was separated from other minor sequence data by data processing such as signal and noise separation and signal screening, and then the main peptide chain was estimated proportion. Comparing the calculated result of this method with the result of mass spectrometry, and using the mixed sample data for verification and analysis. Results The results of this method are basically in line with the actual situation, can be more accurately estimate the proportion of the main peptide chain. Conclusion The method of estimating the proportion of main peptide chain of N-terminal non-uniform single-stranded recombinant protein product by using N-terminal amino acid sequencing data was initially established, which provided assistance for the analysis and research of related proteins.