SBIR-STTR Award

PROGRAM Monoclonal antibodies (mAbs) have become mainstream therapeutic proteins in the biopharmaceutical industry (biopharma), and the demand for efficient analytical tools for their characterization and quality control continues to increase. During manufacturing and storage, cleavage of mAb’s primary structure can occur due to different fragmentation induced mechanisms, and the resulting fragment species can have negative implication for the safety and efficacy of the product. Therefore, fragmentation is a critical quality attribute routinely monitored to assess the purity and integrity of mAbs and mAb-derived biologics from production to commercialization. Capillary electrophoresis with sodium dodecyl sulfate (CE-SDS), which is the capillary analogue of polyacrylamide gel electrophoresis (SDS-PAGE), is widely employed for monitoring mAb fragments during production, formulation, stability, and commercial release. CE-SDS employs SDS to denature proteins and render non-covalently associated subunits separable. The separation is based on electrophoretic migration driven by the surface charge induced by SDS binding, which is proportional to the protein’s molecular weight. In a sieving gel matrix, a size-based separation is achieved since all SDS- protein complexes have similar charge-to-size ratio. CE-SDS provides excellent resolution of fragments, but lack of mass identification through direct coupling to mass spectrometry of the resolved fragments currently creates a major gap in the biopharma. GMJ Technologies (GMJ) is requesting SBIR Phase I funding to develop a micro SDS depletion electrophoresis (?SDE) interface to allow direct coupling of CE-SDS to mass spectrometry (MS). The proposed interface will use free-flow zone electrophoresis mechanism to remove SDS in-line from the CE-SDS peaks of interest prior to electrospray ionization mass spectrometry (ESI-MS). With this innovation, GMJ aims to address a critical need in the characterization and quality control of mAbs from research to commercialization. By eliminating several laborious steps, the proposed device will significantly improve the efficiency in the characterization workflow for mAbs’ fragments, improve the analysis precision and sensitivity, and provide cost-saving benefits for biopharma researchers.

Public Health Relevance Statement:
PROGRAM NARRATIVE This Small Business Innovation Research Phase I project will produce a novel technology for direct mass spectrometry identification of monoclonal antibodies fragments resolved by capillary electrophoresis with sodium dodecyl sulfate, which is crucial to meeting regulatory requirements for biopharmaceutical manufacturers. This will address a major gap in monitoring fragment impurities in biopharmaceutical products, and provide a significant value to the development of therapeutics derived from monoclonal antibodies.

Project Terms:
Angiotensin I; Angiotensins; Clinical Treatment Moab; mAbs; monoclonal Abs; Monoclonal Antibodies; Back; Dorsum; Biological Products; Biologic Products; Biological Agent; biologics; biopharmaceutical; biotherapeutic agent; Biotechnology; Biotech; Buffers; Blood capillaries; capillary; carbonate dehydratase; Carbonate hydro-lyase; Carbonic Anhydrases; Charge; Liquid Chromatography; Digestion; Electrodes; Electrophoresis; Electrophoretic Fractionation; Polyacrylamide Gel Electrophoresis; Polyacrylamide Gel Fractionation; Fractionation; Chemical Fractionation; FRACN; Fractionation Radiotherapy; Gel; Glass; Goals; Gold; Human; Modern Man; Immunoglobulin Fragments; Antibody Fragments; Immunoglobulins; Immune Globulins; Light; Photoradiation; Mainstreaming; Educational Mainstreaming; achievement Mainstream Education; Molecular Weight; Muramidase; Lysozyme; N-Acetylmuramide Glycanhydrolase; Peptidoglycan N-acetylmuramoylhydrolase; Myoglobin; Noise; Ovalbumin; Peptide Mapping; Peptide Fingerprinting; Production; Proteins; Quality Control; Research; research and development; Development and Research; R & D; R&D; Research Personnel; Investigators; Researchers; Safety; Bovine Serum Albumin; Signal Transduction; Cell Communication and Signaling; Cell Signaling; Intracellular Communication and Signaling; Signal Transduction Systems; Signaling; biological signal transduction; Silicon; Si element; Sodium Dodecyl Sulfate; Sodium Lauryl Sulfate; Sulfuric acid monododecyl ester sodium salt; Mass Spectrum Analysis; Mass Photometry/Spectrum Analysis; Mass Spectrometry; Mass Spectroscopy; Mass Spectrum; Mass Spectrum Analyses; Vertebral column; Spinal Column; Spine; backbone; Stress; Technology; Testing; Universities; Washington; Cost Savings; improved; Procedures; Capillary Electrophoresis Fractionation; Capillary Electrophoresis; Surface; Refractory; Phase; Evaluation; analog; Funding; Collaborations; instrument; programs; Pattern; System; interest; meetings; meeting; Spectrometry, Mass, Electrospray Ionization; ESI Mass Spectrometry; ionization; Performance; solute; lithography; disulfide bond reduction; Structure; Exclusion; new technology; novel technologies; Devices; Excision; Abscission; Extirpation; Removal; Surgical Removal; resection; Modeling; Sampling; Proteomics; therapeutic protein; native protein drug; pharmaceutical protein; protein drug agent; protein-based drug; Manufacturer; Size Exclusion Chromatography; Molecular Sieve Chromatography; Molecular Interaction; Binding; protein complex; Incubated; Address; Detection; Resolution; resolutions; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Monitor; Process; Development; developmental; nanofabricate; nanofabrication; Outcome; migration; Coupled; Coupling; innovate; innovative; innovation; analytical tool; commercialization; therapeutic agent development; therapeutic development; Phase I Study; phase 1 study; Industry Standard; Formulation; experiment; experimental research; experiments; experimental study; Computerized Models; computational modeling; computational models; computer based models; computerized modeling; Computer Models; detection limit; biopharmaceutical industry; biopharmaceutical company; manufacture; fabrication