An internal standard indicator for protein analysis by capillary gel electrophoresis

A gel electrophoresis and capillary technology, which is applied in the field of capillary gel electrophoresis and protein analysis, can solve problems such as misjudgment of risk impurities and influence on the calculation of the purity of samples to be tested, and achieve good stability, accurate calculation, and avoid misjudgment Effect

Active Publication Date: 2018-11-06
GENOR BIOPHARMA
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Problems solved by technology

[0005] Since the general internal standard currently used is Beckman Coulter / AB SCIEX's 10KD internal standard, which is a synthetic peptide, there are not only differences between batches, but also it is easy to cover the fragments of the samples to be analyzed signal, which affects the purity calculation of the sample to be tested, and also brings misjudgment of risk impurities

Method used

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  • An internal standard indicator for protein analysis by capillary gel electrophoresis
  • An internal standard indicator for protein analysis by capillary gel electrophoresis
  • An internal standard indicator for protein analysis by capillary gel electrophoresis

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Embodiment

[0034] 1.1 Main instruments: PA800PLUS capillary electrophoresis instrument from Beckman Company.

[0035] 1.2 Experimental materials: The names, article numbers and manufacturers of the experimental materials used in the specific examples are shown in Table 1. The following protein samples in the examples of the present invention refer to anti-VEGF monoclonal antibodies.

[0036] Table 1

[0037]

[0038] Note: The above NA means no article number (because it is configured according to the needs when used) or no remarks; the M in the above 0.1M means mol / L.

[0039] 1.3 Preparation of buffer, protein sample solution and required solution

[0040] (1) Sodium hydroxide with a concentration of 0.1M: Weigh 4.00±0.01g of NaOH, add water to make up to 1000mL, mix well and store at room temperature.

[0041] (2) Hydrochloric acid with a concentration of 0.1M: Measure 9 mL of concentrated hydrochloric acid (HCl mass concentration: 36.5%), add water to make up to 1000 mL, mix we...

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Abstract

The present invention discloses an internal standard indicator for analyzing a protein through capillary gel electrophoresis, and a determination method for determining a protein sample by using the internal standard indicator. According to the determination method, benzoic acid is adopted as an internal standard indicator and a reduction method capillary gel electrophoresis method is used to determine the purity of a protein sample, wherein the protein is preferably an anti-VEGF monoclonal antibody, the determined concentration range of the protein sample is 0.3-2.0 mg / ml, and the preferred concentration of the internal standard indicator benzoic acid is 3 mg / ml. According to the present invention, with the internal standard indicator, the problems of the difference between the internal standard indicator batches and the influence on the purity analysis of the determined sample can be completely solved while the new impurity cannot be introduced, the stability is good; and the internal standard indicator does not easily cover the sample fragment signal of the analyzed protein sample, the purity of the protein sample can be accurately calculated, and the misjudge on the risk impurity is avoided.

Description

technical field [0001] The invention relates to protein analysis and capillary gel electrophoresis (CGE-SDS), in particular to an internal standard indicator applied to capillary gel electrophoresis analysis of proteins and a method for determining protein samples using the internal standard indicator. Background technique [0002] The internal standard (Internal Standard) 10kDa Marker of Beckman Coulter Company (its parent company is AB SCIEX Company) is mainly used as a migration marker in the CGE-SDS (capillary gel electrophoresis) analysis method for Calculate the position of other unknown proteins relative to this protein, so as to estimate the unknown protein. However, in the analysis of known protein samples (such as QC test), since the molecular weight of the protein sample is known, the main function of the 10kD marker is to judge whether the test conditions are acceptable according to its peak time. For QC samples, the stability of the added internal standard and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447G01N33/68
Inventor 顾俊林军
Owner GENOR BIOPHARMA
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