Dr. Rodney Bannwart, a principal scientist at Emery Pharma, discusses LC-MS peptide mapping of biologics. He has extensive experience in LC-MS analysis and has worked on biologic compounds throughout the development process. Peptide mapping involves characterizing sequence coverage, post-translational modifications, disulfide scrambling, glycosylation, and sequence variants in biologics. Biologics, such as monoclonal antibodies, require multiple analytical methods due to their larger molecular weights compared to small molecule pharmaceuticals. LC-MS peptide mapping is the gold standard method for confirming the primary sequence of biologics. The process involves enzymatic digestion of the protein sample and injection into Emery Pharma's high-field Orbitrap UPLC-MS system. Peptides are identified based on observed versus expected molecular masses. The workflow includes denaturation, reduction, alkylation, and digestion of the protein sample. Trypsin or other enzymes are used to generate smaller peptides for chromatography. Achieving 100% sequence coverage is challenging, so complementary hydrolytic enzymes like Glu-C can be employed. The peptide mapping chromatogram shows complex results with numerous peptides, but software analysis and manual confirmation enables their identification. In summary, LC-MS peptide mapping provides valuable information on sequence verification, post-translational modifications, and sequence variants. This method is essential for early-stage identification, comparability, characterization studies, and GMP release in biologics. For further inquiries, contact Emery Pharma at emerypharma.com.
About Emery Pharma:
Emery Pharma is a full-service contract research laboratory, supporting the end-to-end development of small molecule drugs, biologics, biosimilars, gene therapy, botanicals, and more.
Emery Pharma's premium services include analytical, bioanalytical solutions, microbiology & cell biology services, drug and impurity characterization, and general R&D and cGMP/GLP support.
At Emery Pharma, our focus is to find innovative solutions for our Clients’ toughest challenges and tailoring those solutions to fit each Client’s unique needs. Our experienced scientific team will manage the process: from drug discovery, proof-of-concept, data management, data analysis, to gathering insights. The team excels at solving scientific challenges, and addressing any issues early in the process, so Clients don’t experience unnecessary setbacks.
Emery Pharma’s experienced chemists and biologists deploy state-of-the-art technology. Instrumentation ranges from high-resolution mass spec (Orbitrap), Triple quad LC-MS/MS, GC-MS, Preparative HPLC, and 400 MHz multinuclear NMR.
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keywords:
LCMS peptide mapping
Biologics
Dr. Rodney Bannwart
Emery Pharma
Alcon
Novartis
Antibodies
Antibody fragments
Therapeutic proteins
Small molecules
Pfizer
QTOF
Orbitraps
Sequence coverage
Post-translational modifications
Disulfide scrambling
N-glycan release
Molecular weights
Chemical properties
Gold standard method
Enzymatic digestion
High-field Orbitrap UPLC-MS system
Denaturation
Reduction
Alkylation
Trypsin
Hydrolysis
Complementary hydrolytic enzymes
Glu-C
Chromatography
Sequence verification
Nismab monoclonal antibody
Oxidation
Deamidation
Glycosylation
GMP release
Early-stage identification
Comparability
Characterization studies
Emerypharma.com