This practical handbook provides drug discovery teams with a guide to mastering ternary complex analysis using in-solution fluorescence cross-correlation spectroscopy (FCCS).
Designed for researchers working with complex, multi-component systems, it explores how FCCS can directly measure ternary complex formation in solution with single-molecule sensitivity — helping teams move beyond binary affinity measurements and gain clearer insight into cooperativity, stability, hook effects, and mechanism of action.
What’s inside the handbook
- An introduction to the challenges of multi-body binding, including why binary affinity alone can miss promising leads and how cooperativity and the Hook Effect influence ternary complex formation
- A practical explanation of how FCCS measures ternary complexes in solution by tracking the co-diffusion of fluorescently labelled binding partners
- Guidance on using FCCS to generate hook plots and investigate complex formation, cooperativity, stability, dissociation rates, and individual Kd values
- Examples of how FCCS can complement existing drug discovery workflows, from screening and lead triage through to SAR and lead optimisation
- Real-world ternary complex case studies, including targeted protein degradation and therapeutic antibody–HER2 complexes
- A streamlined four-step TrueTernary workflow, covering protein labelling, plate setup, automated measurement on the EI-FLEX Pro, and analysis of ternary complex fractions and hook plots
Download this guide if you want to:
✓ directly measure ternary complex formation in solution rather than infer it from binary interactions
✓ characterise hook effects, cooperativity and complex stability to support more confident lead selection
✓ complement existing screening and biophysical techniques with sensitive, immobilisation-free measurements
✓ establish a practical FCCS workflow for targeted protein degradation, multi-specific biologics and other multi-body drug discovery applications
Tags
Technology
Everything you need to understand and apply single-molecule FRET
This practical guide distils the essential principles, experimental workflows, and analysis strategies behind modern single-molecule Förster Resonance Energy Transfer (smFRET).
Written for researchers at all levels – from first-time users to experienced practitioners – it provides the knowledge needed to generate reliable smFRET data and interpret it with confidence.
What’s inside the handbook
Clear explanations of core concepts, including FRET efficiency, stoichiometry, ALEX, burst analysis, and accurate FRET correction
Step-by-step insights into how confocal smFRET works, how single molecules are detected, and how true single-molecule sensitivity is achieved
Practical examples and case studies showing how smFRET reveals structural heterogeneity, conformational dynamics, and nanoscale distance changes
Comparisons with complementary single-molecule and structural biology techniques, highlighting when and why smFRET is the right tool
Guidance on advanced analysis methods such as E–S plots, burst search strategies, and correction-factor estimation for high-precision distance measurements
Download this guide if you want to:
✓ add a fast, solution-based method that resolves conformational changes at the single-molecule level
✓ complement static structural techniques with dynamic data
✓ gain deeper functional insights and explore new biological questions
