Influence of acylation on the adsorption of GLP-2 to hydrophobic surfaces

Competitive Adsorption of Monoclonal Antibodies and Nonionic Surfactants at Solid Hydrophobic Surfaces

“…Quartz slides from TIRF Technologies (Cary, North Carolina) were cleaned by the procedure adapted from Kern and Puotinen where the surfaces are first immersed in a solution prepared from 25% NH 3 , 30% H 2 O 2 , and H 2 O (1:1:5, by volume) at 80°C for 5 min, rinsed with water, immersed in 30% HCl, 30% H 2 O 2 , and H 2 O (1:1:5, by volume) at 80°C for 5 min, and subsequently rinsed with water and ethanol. The clean quartz surfaces are modified with (3, 3, 3‐trifluoropropyl)chloromethylsilane using vapor deposition as previously described under argon atmosphere . To assure proper and homogenous modification, the contact angles of a sessile drop of ultrapure water on the silanized surfaces were determined.…”

“…The flow cell was fitted into a Spex Fluorolog 3–22 (Jobin Yvon Horiba, Longjumeau Cedex, France). Following a modified protocol from Pinholt et al, a stable baseline was established at a constant flow rate of 4.17 μL/s. Subsequently, a constant wavelength analysis of a concentration range of nonadsorbing external standards of N‐acetyl‐tryptophan was performed, followed by the protein sample (10 mg/mL).…”

“…The clean quartz surfaces are modified with (3, 3, 3-trifluoropropyl)chloromethylsilane using vapor deposition as previously described 35 under argon atmosphere. 36 To assure proper and homogenous modification, the contact angles of a sessile drop of ultrapure water on the silanized surfaces were determined. Contact angles were found to be 89 ± 1 • , measured at three different sites on the surface with a Krüss G2 contact angle measuring system (Krüss GmbH, Hamburg, Germany).…”

Lipidation Effect on Surface Adsorption and Associated Fibrillation of the Model Protein Insulin

“…As a pharmaceutical tool, lipidation of peptides and proteins can be used to increase the circulation time in the bloodstream, and thus increase the therapeutic efficacy of biomolecular compounds [4][5][6] . However, modifying soluble proteins with a hydrophobic moiety such as a lipid tail will inevitably have an effect on the physiochemical properties of the protein, including its physical stability [7][8][9][10][11] . Moreover, protein drugs encounter several surfaces and interfaces during the entire lifetime of the compounds: from production, purification and storage to administration, delivery and in vivo utilization.…”

“…Previously, the lipidated variant of the therapeutic protein glucagon-like peptide-2 (GLP-2) was shown to have high affinity for hydrophobic solid surfaces, where the attached lipid chain increased the amount of GLP-2 adsorbing per unit surface area 11 . In bulk assays, lipidation also affected the fibrillation tendency in a site-specific manner of proteins prone to form amyloid fibrils 23,24 .…”