Background introduction
Monoclonal antibodies prepared by recombinant biotechnology (hereinafter referred to as recombinant monoclonal antibodies) are an important class of drugs in biological products. In recent years, due to its precise biological effects such as targeted killing and neutralization, recombinant monoclonal antibodies have been widely studied and applied in tumors, autoimmune diseases, etc., and in other diseases such as neurology and ophthalmology. Research has also begun to develop gradually. At present, domestically produced recombinant monoclonal antibodies mainly use eukaryotic cell culture expression methods. In process research and quality research, pharmaceutical companies pay more attention to the structural research and quality control of the main active ingredients, but the impurities in the product composition and There are few studies or related concerns about related substances. Considering that these impurities or related substances are important components in recombinant monoclonal antibodies, it will affect the quality of the products, the safety and effectiveness of clinical applications.
The method mainly uses the CAPEL-205 high performance capillary electrophoresis apparatus to measure the related impurities of the products in the recombinant therapeutic monoclonal antibody (immunoglobulin G, IgG), and refers to the American Pharmacopoeia Chapter 129 (Analytical Procedures for Recombinant Therapeutic Monoclonal Antibodies, Capillary The method in SDS electrophoresis can also refer to the method in the Chinese Pharmacopoeia 2015 edition of the three three-part general rule 3127 (Molecular Size Molecular Variant Determination).
1 test conditions
USP129. Reduction conditions
BGE: According to the gel conditions in USP 129
Capillary electrophoresis: Ltot = 30 cm, ID = 50 μm
Sample injection: -5 kV x 20 sec
Voltage: -15 kV
Room temperature: 25 ОС
Detection wavelength: 220 nm
Sample: Recombinant IgG preparation
IS – internal standard
LC – light chain
NG – non-glycosylated heavy chain
HC – heavy chain
USP129. Non-reducing conditions
BGE: According to the gel conditions in USP 129
Capillary electrophoresis: Ltot = 30 cm, ID = 50 μm
Sample injection: -5 kV x 20 sec
Voltage: -15 kV
Room temperature: 25 ОС
Detection wavelength: 220 nm
Sample: Recombinant IgG preparation
IS - internal standard
F1-F8 - IgG related impurities (fragment)
IgG - intact antibody
2 test results
Figure 1 Detection chromatogram under reducing conditions
Figure 2 Detection chromatogram under non-reducing conditions
3 conclusions
The CAPEL-205 high performance capillary electrophoresis system can be used to determine the related impurities in the recombinant therapeutic monoclonal antibody (immunoglobulin G, IgG), and the effective separation of related impurities in the IgG product can be achieved. The method is accurate, sensitive and reproducible. Good sex, can meet the daily analysis requirements.
4 configuration list
Instrument configuration | CAPEL-205 capillary electrophoresis system with laptop with WINDOWS® XP/ 7/8/10 operating system |
ELFORUN dedicated software workstation |
Advantages of Capel-205 Capillary Electrophoresis System
By applying a high voltage electric field, the difference in the migration speed of the components of the capillary liquid sample is based. Application of UV absorption detection technology to quantitatively detect the separated components by using a high-precision optical system at the capillary outlet
Light source: high-intensity xenon lamp, optimized optical design to ensure low detection limit
Can realize a variety of different analytical methods and separation modes, capillary zone electrophoresis, capillary micellar electrokinetic chromatography, capillary isoelectric aggregation, capillary gel electrophoresis, capillary isotachophoresis
Fully automatic injection, optional injection and electric pressure injection. Pressure injection, step value 1 mbar, up to 99 mbar; electric injection, voltage range: step value 1kV, 1-25kV adjustable
Capillary cooling: constant temperature water cooling system, ambient temperature 5-50 °C adjustable; constant temperature water cooling to ensure more precise and accurate temperature control, improve analysis results (relative to air cooling, better cooling effect / lower cost than liquid cooling)
Standard centrifuge tube 1.5mL (Eppendorf type) for all sample analysis
Extended fully automatic injector with up to 59 injection positions, allowing automatic overnight analysis
Unique fully automatic open injection design to avoid volatilization and cross-contamination
The new high pressure module provides ±30kV pressure for superior analytical efficiency and fast analysis time
Using flow potential control technology to greatly improve reproducibility, especially for analyzing complex samples
Source: LUMEX Analytical Instruments
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