Peptide mapping method

An analysis method and peptide mapping technology, applied in the field of peptide analysis, can solve the problem of difficult detection of hydrophilic peptides, and achieve the effect of good separation effect and unified method.

Active Publication Date: 2022-04-19
深圳汉腾生物科技有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Based on this, it is necessary to provide a peptide map analysis method for the difficulty of detecting hydrophilic peptides in traditional liquid chromatography-mass spectrometry separation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0070] In this example, a flexible linker (Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Gly-Ser-Ala-Glu-Ser-Lys) was synthesized. The peptide segment was mixed with bovine serum albumin as the test sample, and the effect of the peptide map analysis method in the present invention on the separation and identification of the hydrophilic target peptide segment in the protein sample was verified.

[0071] (1) Experimental reagent consumables and instruments are shown in Table 2 and Table 3.

[0072] Table 2 Experiment reagent consumables

[0073]

[0074] Table 3 Experimental Instruments

[0075] equipment name Instrument model UPLC Waters-H-class plus Constant temperature metal bath Hangzhou Miou Instrument Co., Ltd. DTK200-4 Vortex shaker Hangzhou Miou Instrument Co., Ltd. MIX-25P High speed refrigerated centrifuge Thermo-LEGEND MICRO 21R centrifuge Thermo-ST16R

[0076] (2) Reagent preparation

[0077] 1×PBS so...

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Abstract

The invention discloses a peptide map analysis method, comprising the following steps: performing denatured enzymolysis on a sample to obtain polypeptide fragments; using UPLC-MS to detect the polypeptide fragments, and analyzing the polypeptide according to the collected peptide map data Fragments are assigned; wherein, the mobile phase of the UPLC-MS includes mobile phase A and mobile phase B, the mobile phase A is formic acid aqueous solution, and the mobile phase B is formic acid acetonitrile solution; the UPLC-MS uses ACQUITY UPLC HSS T3 column for gradient elution.

Description

technical field [0001] The invention relates to the technical field of polypeptide analysis, in particular to a peptide map analysis method. Background technique [0002] Peptide map analysis can provide rich structural information in the confirmation of primary structure, and the peptide map analysis method based on liquid chromatography-mass spectrometry is widely used in the quality control of biopharmaceuticals because it can match with the theoretical amino acid sequence. Applications. Reverse-phase chromatography (RP-HPLC) has become the main tool for the separation and identification of peptides. [0003] C18 chromatographic column is widely used in peptide map analysis due to its unique resolution. However, some hydrophilic peptides produced by restriction enzyme digestion are difficult to be monitored on C18 column. In addition, in order to obtain a good peak shape, trifluoroacetic acid (TFA), a strong ion-pair reagent that can improve the peak shape, is usually a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/72
CPCG01N30/72
Inventor 欧阳玥阮路孟凡菊陈炯高原
Owner 深圳汉腾生物科技有限公司
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