Monoclonal antibodies (mAbs) play a major role in the treatment of cancer, infectious diseases, allergies, inflammation, auto-immune diseases and other human diseases. Antibody-based therapeutics constitute one of the fastest growing classes of pharmaceutical products. Antibodies are characterized by large, complex and heterogenous. To ensure regulatory compliance, extensive analytical characterization is required to identify and control key attributes that affect the safety, stability and efficacy of antibodies. With more than 20 years of experience and continuous innovation, Creative Proteomics has established the industry's most advanced system for measuring and understanding the antibodies. Here, we apply Liquid Chromatography-Mass Spectrometry (LC-MS), a powerful analytical technology, to analyze a wide range of attributes of antibodies, such as sequence integrity, post-translational modifications (PTMs), and epitope identification.
LC-MS— a powerful analytical tool for analyzing antibody-based therapeutics
The wide availability of LC columns, well-established analytical methods and the robustness of the system have made LC the standard introduction system for mass spectrometric characterization of protein therapeutics. To obtain structural domain-specific information and effectively characterize protein therapeutics, monoclonal antibodies are reduced to heavy and light chain components or enzymatically cleaved into smaller fractions or peptides. LC is widely used to separate antibody fragments, and mass spectrometry (MS) is used to identify antibody fragments. LC-MS is used to characterize antibody features including intact molecular weight determination, PTM analysis, higher order structure analysis, and epitope identification.
LC-MS analysis of intact antibodies
For intact antibody analysis, we have developed a highly sensitive and stable LC/MS workflow. Our workflow is based on optimized sample preparation, improved LC-MS instrumentation, and advanced analytical software for sensitive detection and characterization of natural intact macromolecular protein complexes. In general, our service consists of five major steps, including experimental design, sample preparation, LC-MS analysis, data processing, and result interpretation and delivery report.
LC-MS has the versatility to monitor a wide range of antibody properties, keeping pace with today's innovations in biotherapeutic design. To achieve sensitive and accurate quantification by LC-MS, efficient and reproducible sample preparation is essential. The optimization of sample preparation mainly focuses on sample purification and enrichment, stable and efficient recovery of antibodies and peptides. LC-MS analysis after affinity enrichment and stable and high recovery of peptides from target antibodies by enzymatic digestion are effective ways to improve sensitivity.
Our service can help researcher investigate the following lists, but are not limited to:
- Confirmation of primary sequence
- Intact molecular weight determination
- A variety of PTM analysis
- Epitope identification
Why choose us?
- A highly sensitive and stable LC/MS workflow
- Sensitivity and specificity of intact protein bioanalysis
- High throughput and cost-effective analysis of intact antibodies
- Obtain high quality complete mAb data for mAb identification and relative glycoform quantification
LC-MS analysis has become an essential tool for the identification, characterization and quantification of intact mAbs and similar high-molecular-weight proteins. If you are interested in our service, please contact us for more details.
Reference
- Kang, Lijuan, Naidong Weng, and Wenying Jian. "LC-MS bioanalysis of intact proteins and peptides." Biomedical chromatography 34.1 (2020): e4633.
