张竞1,安进城2,李丹丹1,周志艳1,任新桐1,邹卫3▲.液质联用法测定重组人脑利钠肽的质量肽图[J].中国医药科学,2025,(5):17-20转66 基金项目:中央引导地方科技发展资金项目(226Z2404G) |
液质联用法测定重组人脑利钠肽的质量肽图 |
Determination of mass peptide map of recombinant human brain natriuretic peptide by liquid chromatography-mass spectrometry |
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DOI: |
中文关键词: 液质联用;肽图;重组人脑利钠肽;心力衰竭 |
英文关键词:Liquid chromatography-mass spectrometry; Peptide map; Recombinant human brain natriuretic peptide; Cardiac failure |
作者 | 单位 | 张竞1,安进城2,李丹丹1,周志艳1,任新桐1,邹卫3▲ | 1.河北冀兆联科技发展有限责任公司研发中心,河北石家庄 050051;2.石家庄医学高等专科学校冀联校区医院中医科,河北石家庄 050057;3.河北科技大学食品与生物学院,河北石家庄 050018 |
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中文摘要: |
[摘要]目的 建立重组人脑利钠肽(rhBNP)的肽图分析方法,同时绘制其质量肽图并鉴定二硫键位点。方法 摸索合适的酶切条件,用液质联用的方法绘制质量肽图和定位二硫键,并进行重复性、精密度和耐用性相关验证。结果 rhBNP经还原及烷基化后进行酶切,胰蛋白酶与蛋白质量比为1∶100时,可以得到稳定的肽图谱。通过分析理论结果,确认二硫键位置。方法学验证结果表明,该方法重复性、精密度及耐用性良好。对连续3批生产的中试产品进行肽图分析,供试品与对照品的肽图谱完全一致。结论 本研究建立的肽图分析方法可应用于rhBNP的鉴别和质量控制。 |
英文摘要: |
[Abstract] Objective To establish a peptide map analysis method for recombinant human brain natriuretic peptide (rhBNP), simultaneously draw its mass peptide map and identify disulfide bond sites. Methods The suitable conditions of enzyme digestion were explored, the mass peptide map was drawn, and disulfide bonds were located according to the liquid chromatography-mass spectrometry method, and the related repeatability, precision and durability were verified. Results After reduction and alkylation, the rhBNP was digested by enzyme. When the ratio of trypsin to protein was 1∶100, a stable peptide map could be obtained. By analyzing the theoretical results, the locations of disulfide bonds were confirmed. The results of methodology verification showed that the method had good repeatability, precision and durability. The peptide maps of the pilot products produced in three consecutive batches were analyzed, and the peptide maps of the test products and the reference products were completely consistent. Conclusion The peptide map analysis method established in this research can be applied to the identification and quality control of rhBNP. |
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