Immunopurification of proteins containing TG-catalyzed ε- γ-glutamyl) lysine bonds was performed using 81D4 Mab pre-bound to sepharose beads using a protocol developed by CovalAb (Lyon, France) and as previously described (31).
In equilibration section, a position-restrained molecular dynamics with the linear constraint algorithm for all bonds was performed using NVT equilibration, Berendsen weak thermal coupling method, and Particle Mesh Ewald method.
When the binding state of any bond (open versus closed) has undergone a change, an update of the force and surface separation at all bonds is performed using the associated elastic Green's function, and the results are used to determine the subsequent events.
Hydrogen-bond analysis was performed using a maximum hydrogen-acceptor distance of 0.25 nm and a minimum donor-hydrogen-acceptor angle of 135° as the geometric criteria.
When the disulfide bonds prediction was performed using DiANNA, which is recent state-of-the-art web-based software determining the cysteine oxidation state and disulfide connectivity of a protein [ 25], the result showed that 4 of 5 cysteine residues might be involved in the formation of two intramolecular disulfide bonds (Cys330-Cyss475 and Cys376-Cys520).
The alumina and prosthodontic composite's bonding strength, so-called shear bond strength test was performed using a Howden Universal Testing Machine (Leamington Spa, England) running at a cross-head speed of 0.5 mm/minute.
Transient liquid phase (TLP) bonding of Al2O3p/Al was performed using Cu, Al Cu and Cu Ti system interlayers.
The push-out bond strength test was performed using a universal testing machine.
The push-out bond strength test was performed using a universal testing machine (model 4411; Instron Corp., Canton, MA, USA).
The microshear bond strength (μSBS) test was performed using a universal testing machine (EMIC DL-500).
