The therapeutic efficacy of antibody drugs might be hampered by their large molecular masses and complex hetero-tetrameric structures. To address these issues and enhance the overall therapeutic efficiency, one of the essential tasks in preclinical drug discovery is to characterize the important parameters about antibody homogeneity. Creative Biolabs offers a range of tailored solutions towards antibody homogeneity and makes the best understanding of interested antibody drug properties.
Antibody Homogeneity Analysis
Since the use of antibodies in therapeutic medicine was first approved by the US FDA in 1980s, antibody products have become one of the main drivers of the growing biologics drug market. The rapid growth in demand for antibody production has been well accommodated by the industrialization of antibody manufacturing and analysis. Antibody homogeneity analysis services at Creative Biolabs include:
| Service Category | Service | Method |
|---|---|---|
| Antibody Fragment Analysis | Antibody Purity Analysis | SDS-PAGE, Capillary Electrophoresis (CE) |
| Antibody Polymer Analysis | Size Exclusion Chromatography (SEC) | |
| Antibody Fragment Analysis | Reversed-phase (RP) Chromatography | |
| Antibody Isoform Analysis | Capillary Isoelectric Focusing (CEIF), MALDI, ESI-MS, | |
| Antibody Host Cell Protein (HCP) Analysis | ELISA, WB, Mass Spectrometry (MS) |
Antibody purity analysis is critical to successful development of antibody biopharmaceuticals. A common technology for antibody purity analysis is sodium-dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The detergent nature of SDS denatures proteins by disrupting their noncovalent bonds, thereby simplifying the molecular structure. The final separation of antibody depends entirely on differences in the relative molecular mass of their denatured polypeptides.
Because of its automated, quantitative nature, capillary electrophoresis (CE) technology is widely used for antibody purity analysis as well. CE provides high-resolution, quantitative analysis relative to SDS-PAGE.
Size exclusion chromatography (SEC) is a chromatographic method that separates molecules based on their particle size. It is usually applied to large molecules or macromolecular complexes such as proteins and polymers.
Reversed-phase chromatography is often and favored used for both peptide and protein separations due to its superior resolution, good repeatability and the compatibility of eluents with mass spectrometry (MS) analysis. The pharmaceutical industry regularly employs reversed-phase high-performance liquid chromatography (RP-HPLC) to qualify drugs before their release.
Antibody isoforms can share a high percentage of amino acid sequence homology and dramatically differ in their concentration, the task for accuracy and efficiency in methodology and instrumentation is challenging. Creative Biolabs provides antibody isoform analysis services by different technique as described in the table.
Expression systems used in the generation of antibody drugs contain many proteins. These proteins, termed host cell proteins (HCPs), have to be removed during antibody purification. Any residual HCP, even low levels, can potentially compromise the safety and efficacy of antibody drugs. Polyclonal antibodies are the main tools for detecting and monitoring HCPs. The most common methods for testing for HCP are enzyme-linked immunosorbent assay (ELISA), 1-D or 2-D electrophoresis followed by western blotting, and MS-related techniques.
Fig.1 Schematic diagram of residual HCPs in biopharmaceuticals. (Huang, 2017)
A comprehensive characterization of antibody structure is required for the final biopharmaceutical product approval and subsequent manufacturing process. Creative Biolabs is one of the well-recognized experts who is professional in antibody homogeneity analysis for a broad range of project objectives. If you're doing research on drug discovery and you need our help, please don’t hesitate to contact us.
Reference
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