De novo sequencing method based on chemically modified and isotopically labeled polypeptide amino acid sequences
CN106645437B · Dalian Institute of Chemical Physics · priority 2015
| Patent | CN106645437B ↗ |
| Status | Active |
| Priority date | 2015-10-30 |
| Filed | 2015-10-30 |
| Published (granted) | 2019-06-11 |
| Assignee | Dalian Institute of Chemical Physics |
| Also published as | CN106645437A |
Abstract
Translated from Chinese 本发明涉及一种基于化学修饰改性和同位素标记的多肽氨基酸序列从头测序方法。利用N端和C端标记后的多肽分子质量以及保留时间的相关性，将不同标记的相同多肽的二级质谱图进行关联。利用不同标记样品中多肽在质谱碎裂过程中形成的N端和C端碎片离子对，对多肽氨基酸序列进行从头测序。本发明通过对多肽使用带有正电荷的试剂进行修饰改性，有利于多肽在质谱中碎裂时形成丰富的碎片离子；同时，通过使用含有不同轻重同位素的试剂进行标记，多肽在质谱中碎裂时形成成对存在的碎片离子，易于分辨，从而降低干扰信号的影响，提高碎片离子选择特异性以及从头测序的速度；利用N端和C端碎片离子的互补性，提高多肽测序准确度和效率。 The present invention relates to a de novo sequencing method of polypeptide amino acid sequence based on chemical modification modification and isotope labeling. Using the correlation between the molecular mass and retention time of the labeled peptides at the N-terminal and C-terminal ends, the MS spectra of the same polypeptides with different labels were correlated. De novo sequencing of the amino acid sequence of the polypeptide was performed using the N-terminal and C-terminal fragment ion pairs formed during the fragmentation of the peptides in different labeled samples by mass spectrometry. In the present invention, the polypeptide is modified and modified by using a reagent with a positive charge, which is conducive to the formation of abundant fragment ions when the polypeptide is fragmented in mass spectrometry; Fragment ions are formed in pairs during splitting, which are easy to distinguish, thereby reducing the influence of interference signals, improving the selection specificity of fragment ions and the speed of de novo sequencing; using the complementarity of N-terminal and C-terminal fragment ions to improve the accuracy of peptide sequencing and efficiency.
Why it is here
Dalian Institute of Chemical Physics: de novo sequencing with chemical modification and light/heavy isotope labelling of peptide N-termini.
Inventors (5)
张丽华, 单亦初, 张珅, 陈玲凡, 张玉奎
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Catalogued papers it cites (1)
- Quantitation and Facilitated de Novo Sequencing of Proteins by Isotopic N-Terminal Labeling of Peptides with a Fragmentation-Directing Moiety (2000) added by the examiner
Matched from the patent’s 3 literature citations by the rules the paper citation graph uses.
Catalogued patents it cites (2)
- US6829539B2 Methods for quantification and de novo polypeptide sequencing by mass spectrometry
- US7723917B2 Method of de novo sequencing of peptide using the MALDI mass spectrometry, method and apparatus for preparing sample for MALDI mass spectrometry
Cited by (4 patent families, 4 organisations)
- 中国科学院大连化学物理研究所 academic: CN112986570B
- 五邑大学 academic: CN114596912B
- 江汉大学 academic: CN114487198B
- 清华大学 academic: CN112147185B
Every family, of any applicant, that cites this one, as Google Patents lists them; an assignee is given its English name.