Engineered antibodies

Unlocking expression and affinity insights pre-purification
I'm Interested
lipid nanoparticles
LNPs
LNP 
lipid nanoparticle

What does FIDA bring to research on engineered antibodies?

01

Pre-purification characterisation of antibodies, nanobodies and bispecifics

  • FIDA users identify single best clone, saving time and sample material
  • Characterisation directly in fermentation media results in a streamlined workflow and early-stage exclusion of non-functional clones
02

Quantity and quality in a single assay

  • Simultaneous expression and functional assessment based on quantity and affinity measurement (Kd) in cell supernatant and fermentation media
  • Characterise absolute affinities (Kd), size (Rh) of complexes
  • 8 QC parameters for an in-depth characterisation
03

Reduced probability of false positive results*

  • *In comparison to ELISA-based technologies
  • Thanks to first principle, direct measurement.
  • Save time thanks to a bulletproof assay
04

Boosted efficiency

  • Characterise all parameters of the system in a single experiment
  • High throughput autosampler (2x50 vials or 96 well format) saves time
  • Quick verification for binary and ternary complex formation
  • Use a single technology for pre-purification clone selection and in-depth post-purification characterisation
  • Choose the of most efficient clones using combined affinity and expression level measurements
05

Save sample material

  • Typical sample usage: 4μL analyte (Protein of Interest), 40 nL indicator (a ligand to Protein of Interest)
  • Unpurified and with no buffer constraints
  • 120 nL indicator-sample used per triplicate measurement

Rapid confirmation of the binary and ternary complex formation in bispecific antibodies research

Step 1

Fida measures the hydrodynamic radius (Rh) of one of the antigens (fluorescently labelled, i.e. the “indicator”).

Step 2

The absolute size of the complex (Rh) serves as a confirmation of complex formation.

Molecular size measurement. Ternary and binary complex formation. Bispecific antibody

Nanobody clone selection and in depth characterisation

01

Initial Processing

  • Immaturazition
  • Lymphosites & LHH Isolation
  • Clone in Phages & Culture in E-Coli
  • Biopanning

Nanobody clone selection and in depth characterisation

02

FIDA Clone Selection

Optimal Clone Selection based on Affinity (Kd) & Expression Levels, measured in a single assay

FIDA users perform assessment of functional performance (affinity) together with assessment of expression level. 

Having two measurements at the same time increases their ability to select the best protein candidates on the first attempt.

Nanobody clone selection and in depth characterisation

03

Single Best Clone Purification

This allows FIDA users to avoid unnecessary purification of poorly performing clones or nanobodies, saving time and sample material.

Nanobody clone selection and in depth characterisation

04

In-depth 
characterisation

After FIDA users select and purify the single optimal clone, they adopt FIDA for in-depth characterisation of absolute affinities (Kd) and size (Rh).

Common challenges in research on engineered antibodies

Challenge
Clone efficacy and stability
Affinity and expression levels measurements delivered in a single assay, before purification stage allow for efficient clone selection, process optimisation and sample savings. On top of that a simultaneous measurement of extent of protein aggregation or misfolding, allows for concurrent quality control.
Protein stability
FIDA delivers 8 QC parameters, such as aggregation, size and polydispersity, which help to assess protein stability and optimise production.
Protein specificity
FIDA analyzes the interaction with target antigens in solution. This provides information on the binding affinity and specificity of nanobodies, which is later used for activity optimization and reduction of the off-target effects.
Screening
With its low sample consumption, FIDA can be used as a tool to screen libraries of nanobodies for their ability to bind specific antigens.
Membrane protein Fidabio

Are you producing nanobodies to immobilise membrane proteins?

Read about the use of FIDA in membrane protein research.
cryo-em

Are you producing nanobodies to act as cryo-EM chaperones?

Read about the use of FIDA in membrane protein research.

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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