Capillary Isoelectric Focusing Method for Rapid Determination of Components in Protein Mixtures

An isoelectric focusing and mixture technology, which is applied in the field of electric focusing, can solve the problems of unreported drug composition and content analysis of composite antibodies, and achieve the effects of shortening sample consumption, strong versatility and shortening test cycle.

Active Publication Date: 2018-03-06
WUXI BIOLOGICS CO LTD +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Although capillary isoelectric focusing has been widely used in the quality analysis of monoclonal antibodies, its application to the composition and content analysis of compound antibody drugs has not been reported

Method used

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  • Capillary Isoelectric Focusing Method for Rapid Determination of Components in Protein Mixtures
  • Capillary Isoelectric Focusing Method for Rapid Determination of Components in Protein Mixtures
  • Capillary Isoelectric Focusing Method for Rapid Determination of Components in Protein Mixtures

Examples

Experimental program
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Embodiment 1

[0036] Example 1 Determination of the relative content of two monoclonal antibodies in a mixed sample by capillary isoelectric focusing

[0037] 1. Experimental conditions

[0038] Prepare the capillary isoelectric focusing sample solution according to the composition in Table 1, and place it in the autosampler of the ProteinSimple iCE280 capillary isoelectric focusing instrument, and use the fluorine-coated capillary assembly (ProteinSimple / 252701, focusing and detection interval length is 5cm) for analysis , 2000mBar sample for 100s, 1.5kV pre-focusing at room temperature for 1min, 3kV focusing for 8min, detection at 280nm, using ChromPerfect software to integrate the spectrum. The integration parameters are shown in Table 2.

[0039] At the same time, the single protein components in the mixed protein sample are also subjected to isoelectric focusing under the same conditions to obtain corresponding isoelectric focusing spectra. The dosage of single protein component isoelectric ...

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Abstract

The invention discloses a capillary isoelectric focusing method for rapidly determining the content of each component in a protein mixture, comprising: 1) mixing an amphoteric carrier, an additive, an isoelectric point marker and a protein mixture sample to be analyzed to form a mixed 2) Load the mixed solution on a capillary column, focus and detect it to obtain an isoelectric focusing spectrum; 3) compare the isoelectric focusing spectrum in step 2) with the single protein components in the protein mixture sample to be analyzed, etc. Compare the electrofocusing spectra to determine the single protein components corresponding to different isoelectric point ranges in the isoelectric focusing spectra of step 2); 4) According to the comparison results of step 3) and the isoelectric focusing spectra of step 2) Map information to determine the content of each component in the protein mixture sample to be analyzed. The invention has the advantages of strong versatility, short analysis period, simple data processing and the like, and is suitable for the production process development and quality control of the protein mixture.

Description

Technical field [0001] The invention relates to a method for rapidly determining the content of each component in a protein mixture, in particular to a capillary isoelectric focusing method for rapidly determining the content of each component in a protein mixture. Background technique [0002] Monoclonal antibody drugs have significant advantages in the treatment of tumors and autoimmune diseases, but they have certain limitations for diseases with more complex targets, such as infectious diseases or diseases involving multiple targets in the body. [0003] Antibody mixture or antibody cocktail is a class of drugs composed of two or more monoclonal antibodies. It consists of a series of confirmed and well-characterized monoclonal compound antibody drugs. These monoclonal antibodies are in sequence, Both the target and the binding activity have sufficient diversity, which can simulate a series of antibodies produced by the human immune system, and thus has obvious advantages in the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/447
Inventor 郭建平黄哲任文霞赵国凤孟斐阙红周伟昌陈智胜
Owner WUXI BIOLOGICS CO LTD
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