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Online alkylation and HPLC chain dismounting method for analyzing N-terminal sequences of human insulin and analogues thereof

A terminal sequence, human insulin technology, applied in the direction of analyzing materials, material separation, measuring devices, etc., can solve the problem of not being able to determine whether amino acids belong to A chain or B chain, etc., to protect physical health, save time, and avoid excessive contact. Effect

Inactive Publication Date: 2021-03-19
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Different from general protein biotechnology drugs, there are two problems to be solved in the N-terminal amino acid sequence analysis of human insulin and its analogs. The A and B chains must be disassembled and measured separately

Method used

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  • Online alkylation and HPLC chain dismounting method for analyzing N-terminal sequences of human insulin and analogues thereof
  • Online alkylation and HPLC chain dismounting method for analyzing N-terminal sequences of human insulin and analogues thereof

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Embodiment 1

[0029] Determination of the N-terminal amino acid sequence of insulin aspart:

[0030] Insulin aspart is a kind of insulin analogue, which is made by replacing proline at position 28 of human insulin B chain with aspartic acid, so as to change the interaction between molecules and achieve the purpose of quick action.

[0031] Using the method provided by the invention, insulin aspart has successfully realized the splitting of A and B chains ( figure 1 ) and cysteine ​​alkylation modification. Determined by the protein sequence analyzer, the N-terminal partial amino acid sequence analysis results are shown in figure 2 .

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Abstract

The invention discloses an online alkylation and HPLC (High Performance Liquid Chromatography)-based chain dismounting method for analyzing N-terminal sequences of human insulin and analogues thereof,which can realize correct determination of N-terminal amino acid sequences of A and B chains of human insulin and analogues thereof, and is economic, environment-friendly and time-saving.

Description

technical field [0001] The invention belongs to the field of drug analysis, in particular to an online alkylation and HPLC-based double-strand splitting method for the N-terminal sequence analysis of human insulin and its analogs Background technique [0002] The amino acid sequence of proteins and polypeptides, that is, the amino acid sequence, is the basis of their structure and function. Therefore, when developing protein and polypeptide drugs, it is necessary to ensure that the amino acid sequence is correct, which requires sequencing of proteins and polypeptides. N-terminal amino acid sequence analysis based on Edman degradation principle and amino acid sequence analyzer is the most classic protein and peptide sequencing method. [0003] Human insulin for the treatment of diabetes is a widely used biotechnology drug. Human insulin has been modified based on different application purposes, and a series of human insulin analogues have been developed, including insulin asp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/74
CPCG01N30/06G01N30/74
Inventor 方均建董方霆吴胜明李惠马昆鹏王德刚赵俊清
Owner ACADEMY OF MILITARY MEDICAL SCI