Biomolecular condensates

Quantitative data on
droplet count and relative size
I'm Interested

What can FIDA bring to your research on biomolecular condensates?

01

Acquire quantitative data: droplet count, relative size and concentration

  • FIDA users don’t receive yes or no responses. Instead, they receive droplet count and relative size measures, which allows them to fast-track their research through quantitative data
  • Measure light-phase concentration at the various conditions of droplet formation
02

Expand your data generation and understanding

  • Work below, at, and above the critical concentration for phase separation
  • Take affinity measurements even for weak interactions
  • Monitor liquid to solid transitions (aggregation)
  • Track kinetics and droplet maturation
03

Screen and characterise – with one instrument

  • Rapid screening of various conditions by quantification of droplets in walk-away mode
  • Characterisation of interactions through affinity measurements
04

Save your time and energy - reduced manual handling

  • Full automation allows our users to press ‘’go’’ and walk away.
  • High throughput instrument with 2 x 96 well plates or 2 x 50 vials
  • Pre-screen before phase separation
  • No manual data handling - save your energy for what matters the most
05

Lower your sample consumption

  • Only hundred nanoliters per condition in the screening stage - increase efficiency
  • Down to 4 μL analyte, 40 nL indicator for characterisation
06

Take control over environmental variables

  • Full temperature control (5–55°C)
  • 3 temperature measurement points: 2 in the autosampler and 1 in the capillary chamber
  • Physiological conditions
  • No ionic strength constraints
  • High sensitivity offers a wide concentration range
  • Unique technological capability of managing stickiness
  • Freedom of work with no pH constraints
07

Sample control

  • Quantification of aggregates
  • Viscosity measurement as an additional QC readout capillary chamber
  • Polydispersity index (PDI)
  • 8 QC parameters in every datapoint

Quantified characterisation

In FIDA assays, each condensate (droplet) is counted, and its relative size is measured, providing our users with a quantitative readout that they can’t get from any other technology currently available on the market.

Our users receive data on relative sizing and droplet formation kinetics, as well as aberrant droplet aging characterisation. It’s all possible thanks to first principles that FIDA technology is built upon.

sources

Dr. Soumik Ray, the author and proprietor of this work, has generously provided the accompanying images and graphics. Full article of his authorship has been published in Springer Nature Protocols and Method Community (Link). The work featured in these images is part of the research paper entitled 'Capillary flow experiments for thermodynamic and kinetic characterization of protein liquid-liquid phase separation' by Stender, E.G.P., Ray, S., Norrild, R.K. et al., published in Nature Communications, Volume 12, Article number: 7289 (2021). The paper can be accessed at https://doi.org/10.1038/s41467-021-27433-y.

Benefits

Higher throughput than fluorescence microscopy imaging
No need for manual data handling
Users trained within hours
Control over environmental variables

Features

Advanced temperature control throughout the analysis
μL of sample consumption for characterisation
nL of sample consumption for screening
Built-in QC
Dedicated software for liquid-liquid phase separation analysis
Automated workflows

Measures

Accurate determination of dilute phase concentration
Relative droplet size distributions
Kinetics of droplet formation and maturation into amyloid fibrils
Determination of binding affinity between the polypeptide undergoing LLPS and LLPS-modulating compounds

This methodology is very sensitive and useful for assessing binding in a quantitative manner using small amounts of auto-fluorescent proteins. Using this method, we can observe clear changes in apparent hydrodynamic radius (Rh) and in fluorescence signal of EYA1 as a function of increasing concentrations of peptides or of small molecules

Dana Farber Institute
CANCER INSTITUTE

''Fida  Instrument provides a ''ruler'' to measure the length of fibrils''

Stefan Rüdiger Ph.D.
Professor of Protein Chemistry of Disease; Head of Department of Chemistry

''The FIDA is very effective for screening small molecules for solubility and interactions with protein targets. A major advantage has been its ability to clearly show drug binding to difficult-to-work-with targets such as intrinsically disordered proteins and amyloids''

Dr. Alexander Taylor
Research Fellow in Biochemistry/Biophysics

I would say that this instrument is quite useful for core facilities at KI or for Swedish research community in general:

  1. it was possible to measure unlabeled membrane proteins and soluble proteins at low concentrations in small volumes very fast (~6 minutes per measurement).
  2. I also like the temperature control and the autosampler which allowed us to run a lot of samples overnight.
  3. I would say that there is a lot of potential to perform other assays as well, such as the Kd determinations etc. (For this, I use ITC which is quite time-consuming and requires a lot of protein).
  4. In addition, the software and user interface for sample runs and data analysis is quite straightforward and intuitive to use.
Postdoctoral Researcher

"What we value about FIDA is the amount of information we can get from a tiny amount of protein in a single QC run. FIDA technology enables us to understand our proteins better - particularly when they behave in unexpected ways. We chose FIDA because it gives us that vital first result quickly - we don't waste our time optimising an assay that isn't going to work."

Duncan Smith
Lead Scientist

"I am convinced. I have never before been able to detect the ternary construct formation of my construct directly in cell lysate – it took less than two hours."

Thomas Smith
Associate Director, Chemical Biology & Therapeutics

"The Fida 1’s unique capability for measuring binding Kd from weak mM level to strong pM level really enhances our bioanalytical capabilities."

Sherry Zhu
Senior Scientist

"FIDA is an in-solution technique, and thus an ideal binding assay for structurally diverse bsAbs because the analysis does not rely on potentially obstructive surface immobilization and hence it is able to perform characterizations that are unbiased by bsAb format and spatial orientation of the binding domains."

Andreas V. Madsen
PhD Student

"After thorough comparisons, it was clear that the Fida 1, amongst others, presents unique opportunities in analysis of membrane proteins, which is essential to us."

Svend Kjæer
STP Deputy

''The Fida 1 system offers a new approach to detergent screening that has significant advantages allowing essential data to be obtained faster using less material (…) this work presents a new approach to characterize membrane proteins that allows users to reducing costs and time as well as to analyze protein expressed at low levels in a short time thus overcoming any stability issues.''

Isabel Moraes
Principal Research Scientist

"I like the ease of use and the straightforward data analysis and, that experiments can be performed in any type of buffers. We get data, where no other biophysical methods worked before."

Cathrine Birck
Head of Integrated Structural Biology Platform

"The method is easy to setup in any standard laboratory and can be adopted for a high throughput (HTTP) screening, which enabled mechanistic studies of RNA nuclease interactions, as well as efficient guide RNA lead discovery (…) Fida 1 offers straightforward assay development, walk away automation, absolute measurement and ultra low sample consumption."

Denny Truong
Principal Research Associate

"FIDA provides in-depth assessment of activity combined with local and global protein structural changes by measuring the overall hydrodynamic radius of the protein with minimal sample consumption. In addition, it is possible to measure the affinity constant for the analyzed interaction."

Dr. Melanie Hug
Research Associate

"The beauty of Fida 1 is that it allows us to conduct a lot of biophysical measurements in one system using a small amount of reagent and a rapid read-out time which is important for our projects and to our customers. (…) Providing a high-quality, protein characterisation QC package is certainly extending and future proofing our capabilities in biophysical characterisation of the proteins we produce."

Mark Abbott
Managing Director

"The Fida 1 has allowed us to assess the performance of MIPs with a much higher throughput than was previously possible on other platforms. MIPs can be developed in as little as 8 weeks, which is a significant improvement on antibodies, and now they can be fully characterized at an accelerated pace too."

Alan Thomson
CEO

"Fida 1 can be used for full characterization of ternary complex formation for targeted protein degradation. The Fida 1 platform only consumes 39 nL of sample for one measurement and thus allows elaborate condition screening using very small amounts of sample material."

Roman Agafonov
Associate Director of Biochemistry, Biophysics & Structural Biology

"The Fida1 is a very user-friendly system. With the Fida software we can quickly design new methods, set up experiments, and analyze data. By far this is a great instrument and technology that has allowed us to run binding assays for LNP in solution and with flexibility to run in any complex media which we were unable to do using traditional binding methods that required ligand immobilization. The FidaBio customer service is amazing, they are very knowledgeable and responsive whenever I run into issues and need assistance. I highly recommend this instrument for LNP or any other nanoparticle characterization. The system and technology are versatile with applications beyond nanoparticles and is a must in the biophysical tool kit."

Biophysical Analytical Department
Senior Scientist, Analytical Development

"Eventually, with FIDA, we managed to characterise our LNPs. And it was done in less than 2 days. We had for a long time been struggling with SPR."

Chemical Biology and Therapeutics
Associate Director

"We had been looking for ways of quantifying our exosomes, and tested many platform, without success. In less than 2 days FIDA had developed an assay and shown trustworthy quantifications directly in fermentation broth"

Rane Harrison
Director, Analytical Development

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