Predicting the effect of single-point mutations on protein stability or protein-ligand binding is a major challenge in computational biology. Free energy calculations constitute the most rigorous approach to this problem, though the estimation of converged values for amino acid mutations is still challenging. To overcome this limitation, we developed tailored protocols to calculate free energy shifts associated to single point mutations. We herein describe QresFEP, which includes an extension of our early protocols to cover all amino acids mutations, and is based on the newest versions of the OPLS-AA forcefields. QresFEP is implemented in an API framework and the graphic interface QGui, for the MD software Q. The complete protocol is benchmarked in several model systems, optimizing a number of variables concerning sampling and the implementation of Zwanzig’s exponential formula and Bennet’s acceptance ratio methods. QresFEP shows an excellent performance on estimating the hydration free energies of amino acid side chain mimics, included their charged analogs. We also examined its performance on a protein-ligand binding problem of pharmaceutical relevance, the antagonism of neuropeptide Y1 G protein-coupled receptor. Here, the calculations show very good agreement with the experimental effect of 16 mutations on the binding of antagonists BIBP3226, in line with our recent applications in this field. Finally, the characterization of 43 mutations of T4-lysozyme reveals the capacity of our protocol to assess variations of the thermal stability of proteins, achieving a similar performance to alternative FEP approaches. In summary, QresFEP is a robust, versatile and user-friendly computational FEP protocol to examine biochemical effects of single-point mutations with high accuracy.