Influence of acylation on the adsorption of GLP-2 to hydrophobic surfaces.
Acylation of proteins with a fatty acid chain has proven useful for prolonging the plasma half-lives of proteins. In formulation of acylated protein drugs, knowledge about the effect of acylation with fatty acids on the adsorption behaviour of proteins at interfaces will be valuable. The aim of this work was to study the effect of acylation on the adsorption of GLP-2 from aqueous solution to a hydrophobic surface by comparing the adsorption of the 3766 Da GLP-2 with that of a GLP-2 variant acylated with a 16-carbon fatty acid chain through a β-alanine linker. Adsorption of GLP-2 and acylated GLP-2 were studied with isothermal titration calorimetry, fixed-angle optical reflectometry and total internal reflection fluorescence. Furthermore, the effect of acylation of GLP-2 on the secondary structure was studied with Far-UV CD. Acylation was observed to have several effects on the adsorption of GLP-2. Acylation increased the amount of GLP-2 adsorbing per unit surface area and decreased the initial adsorption rate of GLP-2. Finally, acylation increased the strength of the adsorption, as judged by the lower fraction desorbing upon rinsing with buffer.
Department of Pharmaceutics and Analytical Chemistry, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark. Charlottepinholt@gmail.com
Charlotte
C
Pinholt
Sebastian J
SJ
Kapp
Jens T
JT
Bukrinsky
Susanne
S
Hostrup
Sven
S
Frokjaer
Willem
W
Norde
Lene
L
Jorgensen
D000215:Acylation; D000327:Adsorption; D053767:Glucagon-Like Peptide 2 / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D053758:Nanoparticles / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry*
D053767:Glucagon-Like Peptide 2; D011137:Polystyrenes