Johannes Reiß (Düsseldorf, DE; Köln, DE), Katharina Vormann (Düsseldorf, DE; Jülich, DE), Dr. Werner Hoheisel (Köln, DE), Dr. Nils A. Lakomek (Düsseldorf, DE; Jülich, DE), Prof. Dr. Michael C. Hacker (Düsseldorf, DE)
Abstract text (incl. figure legends and references)
Introduction
Antibodies are highly sensitive to aggregation, denaturation and other processes which may negatively affect their quality and safety. Therefore, stabilizers like amino acids, sugars and surfactants are needed.[1] We created a platform of macromolecules which combine these different excipient classes in one molecule.
Objectives
Creating a platform of derivatized amphiphiles which stabilize antibodies via oligovalent interactions of hydroxyl, hydrophilic and hydrophobic functional groups of selected amino acids.
Material & Methods
Anhydride-containing amphiphiles were synthesized by radical polymerization of vinyl myristate, acryloyl morpholine and maleic anhydride in various ratios to adjust anhydride content. These amphiphiles were derivatized with different short di(amino acids) and amino polyols via anhydride-amine conjugation.
Interactions of derivatized amphiphiles and a 15N-labeled model protein Nsp3a Ubl1 from Sars-CoV-2 were investigated by 1H-15N HSQC NMR. Interaction sites of the protein were identified based on an established method.[2]
Results
We were able to create an easily adjustable platform of oligo(amino acid) and polyol derivatized amphiphiles and confirmed their structure via 2D-NMR. First studies with a model protein showed specific hydrophobic as well as ionic interactions.
Conclusion and Outlook
Ongoing studies are focused on amphiphiles derivatized with oligo(amino acids) with other functional side chains and their interactions with the model protein.
References
Jorgensen L et al. Recent trends in stabilising peptides and proteins in pharmaceutical formulation – considerations in the choice of excipients. Expert Opinion on Drug Delivery. November 2009;6(11):1219–30. Serrano P et al. Nuclear Magnetic Resonance Structure of the N-Terminal Domain of Nonstructural Protein 3 from the Severe Acute Respiratory Syndrome Coronavirus. Journal of Virology. November 2007;81(21):12049–60.