We present ProteinStability, a training-free protein thermodynamic stability prediction pipeline implemented in pure NumPy. Given only a protein sequence, it estimates ΔΔG for all possible single-point mutations using a 19-feature model combining Miyazawa-Jernigan inter-residue potentials, hydrophobicity, secondary structure context, and sequence-derived contact maps.
Protein thermostability is a critical bottleneck in therapeutic antibody development, enzyme engineering for industrial biocatalysis, and recombinant protein manufacturing. Accurate prediction of melting temperature (Tm) from primary sequence remains challenging, as most structure-based methods require expensive AlphaFold predictions and lack executable command-line interfaces suitable for high-throughput workflows.
Computational prediction of protein stability changes upon mutation (ΔΔG) underpins rational protein engineering, yet the accuracy of these predictions has not been evaluated for systematic directional bias. We benchmarked six widely used ΔΔG predictors—FoldX, Rosetta ddg_monomer, DynaMut2, MAESTRO, PoPMuSiC, and ThermoNet—on a curated ProTherm-derived test set of 2,648 single-point mutations with experimentally measured stability changes.