Pyrazinamide (PZA) plays a unique role in shortening Mycobacterium tuberculosis (TB) treatment compared to other first-line anti-TB drugs. We optimize PZA encapsulation into poly(lactide-co-glycolide) (PLGA) nanoparticles through Taguchi experimental design in a strategy to improve drug bioavailability, reduce the dosing frequency and improve patient adherence to prescribed therapy, one of the critical obstacles in the control of TB epidemics. PZA-loaded PLGA nanoparticles were prepared by the double emulsion method. The Taguchi method, a statistical design with an orthogonal array, was implemented to optimize the formulation parameters. The solvent type (dichloromethane and ethyl acetate), the PZA/PLGA weight ratio and the organic/aqueous phase volume ratio were chosen as significant parameters affecting the particle size and polydispersity index (PDI). Taguchi method proved to be a quick, valuable tool in optimizing the formulation. The optimized experimental values for the nanoparticle size and PDI were about 170 nm and ˂ 0.1. Optimized PLGA nanoparticles possessed a zeta potential of about −1 mV, an encapsulation efficacy of 7–8% and a drug loading of 3.1%. They are suitable to be tested in vivo for TB treatment.