September 2020

Applications News

Membrane embedded proteins: Ligand-binding characterization of Nanodisc-embedded GPCR (β2AR).

G protein-coupled receptors (GPCRs) are the largest family of membrane proteins. They are involved in many vital physiological cell mechanisms, amongst others hormones signalling, cell cycle regulation, neurotransmitters signalling and smell receptors. GPCRs-targeting drugs represent 27% of the global commercial drugs. Therefore, new technologies in this area will advance new discoveries and ultimately new treatments for GPCR related disorders. In this work, it is shown that Flow-Induced-Dispersion-Analysis (FIDA) can be used to fully characterize the binding between Nanodisc-embedded β2-adrenergic receptor and Nanobody (Nb80). In addition to the ability to analyse the functions of the complex, the built-in quality control validates sample integrity and the platform also provides information on the oligomeric state of the receptor. This application is another example of the accuracy of our capillary-based technology for measuring complex bindings through absolute and accurate size determinations of analytes in a pressure-driven flow system.

Materials & Methods
Fida 1 platform with 480 nm LED fluorescence detection for binding

experiments (Fida Biosystems ApS). FIDA standard capillary (i.d.: 75 µm, LT: 100 cm, Leff: 84 cm). HEPES buffer 10mM pH 7.4, 0.5 mg/ml BSA was used as the working buffer. Nanobody Nb80 was used as the indicator. Nb80 was labeled with an Alexa Fluor® 488 Protein Labeling Kit from ThermoFisher Scientific. NanoDisc-embedded β2-adrenergic receptor (β2AR) was used as the analyte (0-1 µM). Sample analysis was performed by filling the capillary with the analyte, followed by an injection of 39 nL of preincubated indicator+analyte, which was mobilized towards the detector at 400 mbar.

These results show how the Fidabio platform may be used to provide functional characterisation of complex membrane proteins and, in the specific case, determine the activity of GPCR embedded in Nano-Discs, based on in-solution and non-invasive analyses. 

For more information, please refer to our application note on the topic.

Nanodisc-enbedded GPCR
Binding curve between Nb80 and ND-β2AR 
Raw data showing the indicator peak getting wider as the concentration of ND-β2AR increased, due to an increase in apparent size upon binding

Transforming Immunogenicity Assessments of Biopharmaceuticals

Over the past couple of years, assessments of immune responses in numerous patient samples have been conducted using our Fidabio technology. The clear advantages are testing and characterization under physiological relevant conditions as well as absolute and quantitative measurements.

As part of Morten Enghave Pedersen’s industrial PhD project, he has established a new application involving characterization of immunogenicity in patients receiving treatment with the anti-TNF-α drug, adalimumab. In only 2 weeks, he has analyzed more than 1,000 data points, and thereby identified patients that had generated neutralizing antibodies and differentiated these from patients with therapeutically active adalimumab.

Morten is in the process of further expanding this unique application.

The figure below shows the development data of the immunogenicity model.

The interactions between TNF-α and adalimumab were fully characterized using the FIDA 1 instrument. The apparent size of TNF-α-alexa488 (100 nM) was measured in a titration series with adalimumab (0-1000 nM).  Here, he observed the formation of both lower and higher-order TNF-α-adalimumab complexes, due to multiple stoichiometries (seen as a hook effect). The binding isotherm was applied to the data set (solid line), revealing an apparent Kd of 1.52 nM and complex size of 8.9 nm. Finally, size-based characterization of the TNF-α-adalimumab complexes allows for immunogenicity assessments in patients receiving adalimumab treatment, by following the apparent size of TNF-α. Hence, the presence of neutralizing antibodies will be reported as a low apparent size of TNF-α, whereas occurrences of binding antibodies will result in high TNF-α size.

We are looking forward to publishing the entire data set. Please get with us, if in the meantime, you are interested in following the project.


New software features enabling fast and agile amendments of sequences

In our continuous effort to improve our software suite, we have just launched a significant new feature that enables users to set up, edit, and adjust sequences on a single parameter level.

It is easy with the Fidabio Assay Design software to set up your experiments and automatically generate the protocols, the methods and sequences needed to run your analyses, incl. the titration series required to establish your binding curves. From time-to-time however, you may want to adjust specific method and sequence parameters. With the new features, we have made such edits as easy as changing the content of a cell in an excel sheet. In the screenshot below editing is done in the table to the left.


3rd annual TPD summit

Fidabio is attending the 3rd Virtual Targeted Protein Degradation Summit, 13-15/10/20 where you will have the opportunity learn how Fidabio can deliver results when it comes to: 1) the absolute size of the ternary complex and the hook, 2) quantitative determination of binding avidity effects, 3) analysis of positive and negative cooperativity, and 4) seemless integration with FRET screening workflows.

Internal stuff

New Team Members

New Sales Director Germany & Austria

We see a steep increase in customer inquiries from Germany and Austria. To service current and future customers in the territory, we have hired Dr Thomas Bauer-Jazayeri as Director, Germany & Austria. Thomas holds a PhD in immunology from Heidelberg University and has more than ten years of sales experience.

Market Research Assistant

The number of new applications is growing, incl. a comprehensive portfolio of applications enabling fast and precise analysis of oligomeric state and applications within the area of targeted protein degradation. To accelerate the market introduction hereof, we have hired Xanthippi Papadopoulou as Market Research Assistant. Xanthippi is studying Business Administration and Bioentrepreneurship at CBS.

New Technician

To address the increased demand for pilots and CRO projects, we are happy that Jan Gøral has accepted to join us as Laboratory Technician. Jan has worked for several years at Copenhagen University.

Please join me in welcoming all three! We are looking forward to continuing the Fidabio journey with them. 

New Grant to fight Covid-19

The primary objective of this project is to develop a dedicated analytical platform that as an orthogonal technology will accelerate the assessment of essential efficacy and safety issues associated with conditional approval of new vaccines against current and future pandemics.

Fidabio with the novel FIDA (flow-induced-dispersion-analysis) technology will collaborate with Adelis (a new highly sensitive UV detection unit) to address critical characterisation and quantification challenges, which are essential to the assessment of efficacy and safety. It will enable fast generation of highly reliable, supplementary data to accelerate emergency authorisation procedures for approval of vaccines for pandemic viruses’ outbreaks, including expediting research projects for vaccines against the current coronavirus outbreak and future modifications hereof.