Targeted Protein Degradation

In-depth characterisation – within minutes, without a doubt.
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What does FIDA bring to TPD research?

01

Effective degrader design

  • Direct measurement of degrader constructs
  • Ternary complex validation by direct measurement of changes in size
  • Hit-characterisation as follow-up to high throughput screens
  • Affinities and functionality of the degraders characterised in 4 minutes
  • Absolute size measurement of the complexes eliminates second-guessing on Kds of all binding partners, incl. cooperativity and sample integrity
02

Directly in cell lysate

  • Measure interactions with GFP/YFP tagged proteins
  • FIDA users save time by not having to purify their proteins
03

Low sample consumption

  • 〜10x less sample consumed than in most SPR technologies
  • Work directly in unpurified samples
  • Down to 4 μL analyte, 40 nL indicator
  • Recover your sample material
04

Affinity measurement readout even for weak interactions

  • The most sensitive affinity measurement on the market
  • Affinity from high pM to mM Kds
05

Direct and in-vitro detection of ubiquitination

  • Save time and energy otherwise spent on SDS-page and western blot
06

Built-in quality control in every data point to increase data reliability

  • 8 parameters: Absolute size, Polydispersity index, Sample loss, PDB correlator, Aggregation, Viscosity, Stickiness, Labelling quality

Benefits

Determine binding interactions under physiological conditions
Know Kds of all binding partners
Know the direct, absolute size
Clear and easy to interpret data dashboard
Speed to data
Troubleshoot efficiently
On-spot analysis kept in-lab

Features

Directly in cell lysate, independent of buffer conditions
Measure cooperativity, even for weak interactions
Sample integrity control
Low sample usage - 4μL analyte, 40 nL indicator
Dedicated TPD software module
Calibration free assay

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

Full quantitative ternary complex characterisation

FIDA provides our users with a full, quantitative characterisation of the ternary complex and its subunits, including Kds of all binding partners, cooperativity factor and sample integrity control.

All under any buffer condition and in complex media, including cell lysate.

  1. Quantitative determination of ternary complex formation
  2. Determination of all construct components bindings, including cooperativity
  3. Determination of the bound fraction of the protein of interest to the ternary complex

Save months of work – FIDA users meet their goals ahead of their competitors

  1. FIDA’s quality control delivers users clear information about the quality of their samples – allowing for troubleshooting in minutes, not days, and saving our users from many frustrations.
  2. FIDA is already used at the expression and pre-purification stage in order to indicate if degraders are functional. This helps to save our users from expensive late stage attritions.
  3. FIDA assays are ready within a few hours, even in the case of new users.

Explore improved workflow opportunities

Thanks to in-depth characterisation, FIDA narrows down the number of leads identified through high-throughput screening

  1. FIDA is employed in diverse workflow stages –
    before or after purification.
  2. It is used for QC, lead characterisation and dedicated parameter determination.
  3. FIDA workflows are shorter thanks to running assays on site – with FIDA there is no need to send out samples to specialised SPR departments

Stay independent

  1. After a short training by our dedicated FIDA TPD team, users run assays on spot and in their own development lab.
  2. With FIDA, there is no more need to send out the samples to external analysis departments.

Improving drug development productivity

  1. In-depth and rapid quality control delivered by FIDA facilitates and speeds up the evaluation
    process in the hit-to-lead stage of drug
    development.
  2. FIDA users increase their efficiency of screening hits, and reduce iterations thanks to FIDA’s first principle nature, the ability to run under native conditions, and the comprehensive built-in QC.

Targeted Protein Degradation

FIDA

Targeted Protein Degradation

tartgeted protein degradation tdp workflow

Targeting common challenges in research on TPD

TPD Challenge
Identify efficient target protein degradation candidates
Screen libraries of degraders – Thanks to FIDA’s capacity to monitor even minute changes in the size of the target protein as a response to different compounds, FIDA can be used for assessment of degraders’ ability to induce degradation of targeted protein.
Occurrence of ubiquitination
Compared to e.g. SDS-PAGE and western blots, FIDA enables real-time monitoring of the ubiquitination reaction in 15–30 mins
Evaluating selectivity
FIDA can be used to analyse whether your candidates actually measure changes in the size of any protein, including the non-target ones. This can make it a tool to evaluate off-target effects of degraders.
Effective degrader design
The dedicated FIDA ternary complex software module enables users to determine the relevant affinities under diverse conditions. This information facilitates and speeds up the selection of degraders with optimal properties.

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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