Liposome-Protein interactions on the Octet RED96e platform

Protein Intractions

University of Natural Resources and Life Sciences, Vienna – BOKU Core Facilities

Jakob Wallner, BSc, Irene Schaffner, PhD, Karin Kohlweiss, BSc

8-Channel Octet Systems: Octet RED96e

The Challenge

Liposomes are transport structures used inside and among cells, and one popular research application is using liposomes for targeted drug delivery, based on specific binding interactions with disease targets. Hydrophobic, spherical vesicles that are composed of at lipids, liposomes can be difficult to study using traditional techniques like SPR, which rely on microfluidic sample delivery.

The Solution

At the University of Natural Resources and Life Sciences Vienna, Austria, the Core Facility Biomolecular & Cellular Analysis (BmCA) offers state-of-the-art instruments and techniques for characterization of biomolecules and/or biomolecular interactions. Since microfluidics lines can be prone to cross contamination and fouling, Core Facility staff member Jakob Wallner tested the Octet RED96e system, which measures binding affinities without the use of microfluidics, eliminating the risk when studying liposomes. The Octet RED96e provides versatility in experimental design, and enabled Jakob to design and run 32 kinetic measurements of liposome-protein interactions during a single unattended run of less than 30 minutes. Besides providing use of the Octet instrument, BmCA also offers professional training and consultation regarding experimental setup and design, full-services such as measurements of user samples by trained staff, and the generation of experimental reports. Additionally, to the quantification of proteins and the measurement of classical protein-protein interactions, users exploit the Octet RED96e for all kinds of biomolecular interactions including interaction studies with carbohydrates, liposomes, nanodiscs, viruses, virus like particles and whole cells. This expertise is reflected in several publications referenced below.

Non-fluidic bio-layer interferometry (BLI) has rapidly become a standard tool for monitoring almost all biomolecular interactions in a label-free, real-time and high-throughput manner. - Jakob Wallner

List of Publications

Wallner, J; Sissolak, B; Sommeregger, W; Lingg, N; Striedner, G; Vorauer-Uhl, K (2019): Lectin bio-layer interferometry for assessing product quality of Fc- glycosylated immunoglobulin G. Biotechnol Prog. 10:e286

Wallner, J; Lhota, G;Schosserer, M; Vorauer-Uhl, K (2017): An approach for liposome immobilization using sterically stabilized micelles (SSMs) as a precursor for Bio-layer Interferometry-based interaction studies. Colloids and Surfaces B: Biointerfaces. 154:186-194

Nika, L.; Wallner, J; Palmberger, D; Koczka, K; Vorauer-Uhl, K; Grabherr, R (2017): Expression of full-length HER2 protein in Sf9 insect cells and its presentation on the surface of budded virus-like particles. Protein Expr. Purif. 136:27-38

Wallner, J; Kuhleitner, M; Brunner, N; Lhota, G; Vorauer-Uhl, K (2014): Application of the log-normal model for long term high affinity antibody/antigen interactions using Bio-Layer Interferometry. J. MATH. CHEM. 52: 575

Wallner, J; Lhota, G; Jeschek, D; Mader, A; Vorauer-Uhl, K (2013): Application of Bio-Layer Interferometry for the analysis of protein/liposome interactions. J. Pharm. Biomed Anal. 72:150-4

Products Used

The 8-channel Octet systems perform quantitation of 96 samples in 32 minutes and kinetic screening of 64 samples in 1.5 hours. Analysis can be done using a single channel or up to eight channels, enabling flexibility in sample throughput based on need. An optional microplate evaporation cover that minimizes losses in sample volume is available with the Octet RED96e.

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