Creative Proteomics provides you with antibody drug charge variant analysis services through isoelectric focusing or ion exchange chromatography technology platforms to help you understand the changes in protein charge heterogeneity during production and purification.
Antibodies can produce a large number of modifications and degradations during production and storage, such as oxidative deamidation, glycosylation, C-terminal lysine truncation, N-terminal pyroglutamate cyclization, disulfide bond modification, sequence variation, etc. These will change the number or structure of the charge groups and cause the heterogeneity of the antibody's charge distribution, thereby affecting the biological activity, pharmacokinetics, immunogenicity and structural stability of the antibody drug. In addition, it may affect the effectiveness, safety and shelf life of the drug in clinical use. Therefore, the detection of antibody drug charge heterogeneity is also an important indicator of the entire production process.
Technology Platform of Charge Heterogeneity Analysis:
- Gel Isoelectric Focusing Electrophoresis (IEF): The separation of proteins by isoelectric point (PI) is routinely used for fingerprint analysis of charge isoforms of recombinant proteins. Advantages: Visual separation and identification according to the charge state and isoelectric point of the protein. The strips can be cut for sample collection for subsequent analysis. But this method cannot be used for quantitative analysis.
- Capillary isoelectric focusing (CIEF): Separate and detect antibodies based on the difference in isoelectric point (pI). The pI value reflects the uniformity of the charge and spatial conformation of the protein drug. Advantages: Compared with IEF, cIEF has higher resolution, analysis speed, quantitative ability and automation performance. Compared with ion exchange gel chromatography, cIEF analysis is faster.
- Imaged capillary isoelectric focusing (iCIEF): The resolution of this method is comparable to that of traditional isoelectric focusing gel electrophoresis, but it has the advantage of whole column imaging. Automatic sample injection, short analysis time, no need to migrate protein to UV detector, and quantitative detection can be achieved.
- Ion exchange chromatography: including cation exchange chromatography and anion exchange chromatography. Ion exchange chromatography can separate proteins with similar isoelectric points. Advantages: Compared with isoelectric focusing electrophoresis, it can detect the difference in surface charge distribution of protein molecules. It can provide chromatographic separation of protein charge isomers. A variety of cation and anion exchange resins provide a variety of separation options. It can separate and collect fractions for the next analysis (after desalting and buffer replacement, mass spectrometry can be performed).
Creative Proteomics will strictly abide by the FDA, ICH, USP and EP guidelines, and provide you with comprehensive analysis services for the charge heterogeneity of antibody drugs. We will customize an exclusive solution for you according to your samples. We are confident to provide you with technical support related to drug development, production process and other special requirements. Look forward to working with you. If you have other questions, please feel free to contact us.
