title: Thermodynamic and structural characterization of an antibody gel.
abstract: Although extensively studied, protein-protein interactions remain highly elusive and are of increasing interest in drug development. We show the assembly of a monoclonal antibody, using multivalent carboxylate ions, into highly-ordered structures. While the presence and function of similar structures in vivo are not known, the results may present a possible unexplored area of antibody structure-function relationships. Using a variety of tools (e.g., mechanical rheology, electron microscopy, isothermal calorimetry, Fourier transform infrared spectroscopy), we characterized the physical, biochemical, and thermodynamic properties of these structures and found that citrate may interact directly with the amino acid residue histidine, after which the individual protein units assemble into a filamentous network gel exhibiting high elasticity and interfilament interactions. Citrate interacts exothermically with the monoclonal antibody with an association constant that is highly dependent on solution pH and temperature. Secondary structure analysis also reveals involvement of hydrophobic and aromatic residues.
authors: Pharmaceutical Development, Genentech, South San Francisco, CA, USA. esue.osi@gene.com
authors: Osigwe
authors: O
authors: Esue
authors: Anna X
authors: AX
authors: Xie
authors: Tim J
authors: TJ
authors: Kamerzell
authors: Thomas W
authors: TW
authors: Patapoff
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry* / Q000378:metabolism*; D016466:CHO Cells; D002151:Calorimetry / Q000379:methods; D019343:Citric Acid / Q000737:chemistry; D003412:Cricetulus; D005782:Gels / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D046529:Microscopy, Electron, Transmission; D019999:Pharmaceutical Solutions; D017550:Spectroscopy, Fourier Transform Infrared; D013329:Structure-Activity Relationship; D013696:Temperature; D013816:Thermodynamics
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D005782:Gels; D019999:Pharmaceutical Solutions; D019343:Citric Acid
keywords: FTIR; ITC; association; carboxylates; citrate; cryoglobulins; gelation; histidine; mechanical properties; monoclonal antibody; rheology; spectroscopy; structure; viscosity