Demo: Relative Binding Free Energy (RBFE)
RBFE Network Workflow

Ligand-X Demo

Demo: Relative Binding Free Energy (RBFE)

See how Ligand-X compares binding affinities across a series of ligands using network-based alchemical calculations.

Video Walkthrough

Coming Soon: Video walkthrough demonstrating the complete RBFE workflow in Ligand-X.


Data Flow Pipeline

When you submit an RBFE calculation, your data flows through these stages:

1. Ligand Preparation
  • Parse SDF files, add hydrogens
  • Assign AM1-BCC charges using RDKit
2. Ligand Alignment
  • 3D alignment using Kartograf atom mapper
  • Compute RMSD to reference structure
3. Network Planning
  • Build transformation graph (MST/Radial/Maximal)
  • Score edges using LOMAP
4. Optional: Docking Validation
  • Validate ligand poses via AutoDock Vina
  • User confirms poses before proceeding
5. Alchemical Transformations
  • For each edge in network: create hybrid topology (OpenFE)
  • Run complex leg (protein + ligand)
  • Run solvent leg (ligand in water)
  • Compute ΔΔG using MBAR
6. MLE Analysis
  • Maximum Likelihood Estimation to convert relative ΔΔG values to absolute rankings
Results
  • Network graph with ΔΔG for each transformation
  • Ranked ligand list with relative binding affinities
  • Uncertainty estimates for each value
  • Alignment data and atom mapping visualizations

Tools Used

Stage Tool Purpose
Ligand Prep RDKit Molecule parsing, hydrogen addition
Charges Antechamber (AM1-BCC) Partial charge assignment
Atom Mapping Kartograf 3D-aware atom mapping
Network LOMAP Scoring transformations
Simulation OpenMM GPU-accelerated MD
Free Energy OpenFE Alchemical RBFE protocol
Analysis PyMBAR Optimal free energy estimation

Example Output

After calculation completes, you’ll see:

  • Network Viewer: Interactive graph showing all ligand transformations
  • Ranked List: Ligands sorted by predicted binding affinity
  • Uncertainty: Error bars showing confidence in each prediction
  • Convergence: Plots showing how estimates stabilized over simulation time

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