title: Taylor Dispersion Analysis as a promising tool for assessment of peptide-peptide interactions.
abstract: Protein-protein and peptide-peptide (self-)interactions are of key importance in understanding the physiochemical behavior of proteins and peptides in solution. However, due to the small size of peptide molecules, characterization of these interactions is more challenging than for proteins. In this work, we show that protein-protein and peptide-peptide interactions can advantageously be investigated by measurement of the diffusion coefficient using Taylor Dispersion Analysis. Through comparison to Dynamic Light Scattering it was shown that Taylor Dispersion Analysis is well suited for the characterization of protein-protein interactions of solutions of α-lactalbumin and human serum albumin. The peptide-peptide interactions of three selected peptides were then investigated in a concentration range spanning from 0.5mg/ml up to 80mg/ml using Taylor Dispersion Analysis. The peptide-peptide interactions determination indicated that multibody interactions significantly affect the PPIs at concentration levels above 25mg/ml for the two charged peptides. Relative viscosity measurements, performed using the capillary based setup applied for Taylor Dispersion Analysis, showed that the viscosity of the peptide solutions increased with concentration. Our results indicate that a viscosity difference between run buffer and sample in Taylor Dispersion Analysis may result in overestimation of the measured diffusion coefficient. Thus, Taylor Dispersion Analysis provides a practical, but as yet primarily qualitative, approach to assessment of the colloidal stability of both peptide and protein formulations.
authors.affiliation: Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark; Early Stage Development, Ferring Pharmaceuticals A/S, Kay Fiskers Plads 11, DK-2300 Copenhagen S, Denmark. Electronic address: ulrich.hogstedt@ferring.com.
authors.forename: Ulrich B
authors.initials: UB
authors.lastname: Høgstedt
authors.affiliation: Early Stage Development, Ferring Pharmaceuticals A/S, Kay Fiskers Plads 11, DK-2300 Copenhagen S, Denmark; F. Hoffman-La Roche Ltd., Pharmaceuticals Division, Grenzacherstr. 124, CH-4070 Basel, Switzerland. Electronic address: gregoire.schwach@roche.com.
authors.forename: Grégoire
authors.initials: G
authors.lastname: Schwach
authors.affiliation: Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark. Electronic address: marco.vandeweert@sund.ku.dk.
authors.forename: Marco
authors.initials: M
authors.lastname: van de Weert
authors.affiliation: Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark. Electronic address: jesper.ostergaard@sund.ku.dk.
authors.forename: Jesper
authors.initials: J
authors.lastname: Østergaard
mesh_terms: D008956:Models, Chemical; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D014783:Viscosity
chemical_list: D010455:Peptides
keywords: Peptide; Protein; Self-interactions; Taylor Dispersion Analysis; Viscosity