Journal Article

Native high-resolution mass spectrometry analysis of noncovalent protein complexes up to 450 kDa

Hottenstein, Charles S, Szapacs, Matthew E, Kellie, John F, Chen, Zhuo

2020

Overview

The structural and functional versatility of immunoglobulinG (IgG) scaffold of mAbs also facilitates the design of more complex compounds, such as antibody-drug conjugates, Fc-fusion proteins and bispecific antibodies (2). Since the functions of mAb-based biotherapeutics are typically achieved by protein-protein or protein-ligand interactions, direct analysis of such noncovalent interactions has great potential to elucidate the pharmacology and toxicology of mAbs. [...]to obtain the information of noncovalent macromolecular assemblies (such as ligand binding, substrate turnover and subunit stoichiometry), native MS has attracted increasing research interest (11,12). [...]the number of charges associated with the protein is usually reduced compared with intact MS analysis, indicating a more compact and folded structure under the native conformation. Because the integrity of proteins is not disrupted, native MS provides a wealth of information for understanding biological properties of intact protein assemblies under near-physiological conditions. The major requirement for buffer in ESI is volatility, which ensures the analyte completely transferring into the gas phase and reduces the formation of adducts with nonvolatile salts impacting ion signals (16). [...]a compatible buffer is also necessary for native MS. Buffer with strong acidity and significant amount of organic solvent can open the native conformation of protein to a less compacted partially folded form and hereby should be avoided (17).

• Publisher: Newlands Press Ltd,Newlands Press
• Subject: Acidity / Adducts / Antibodies / Antigens / Bispecific antibodies / Data processing / Drug development / Fc receptors / Gas flow / Immunoglobulin G / Ions / Laboratories / Ligands / Macromolecules / Mass spectroscopy / Medical research / Monoclonal antibodies / native mass spectrometry / non-covalent protein complex / Peptides / Pharmaceutical industry / Protein structure / Proteins / Salts / Software / Stoichiometry / Structure-function relationships / target engagement