Green fluorescent protein (GFP) is widely known as a molecule capable of monitoring industrial processes due to its ability to emit fluorescence under a broad temperature and pH range.  Improving GFP purification by a rapid and low cost method that delivers high yields (>70% of the initial fluorescent intensity) of pure protein (> 95%) is necessary to meet the demand for use in applied biotechnology.  This work aims to compare three different pre-packed anion exchange chromatography resins to optimize GFP purification and determine the most appropriate matrix.  GFP (pI = 4.7 - 5.1) was extracted from E. coli DH5-alpha cells by a three-phase-partitioning method and 0.5 mL aliquots were loaded onto 1 mL HiTrap ion exchanger chromatography columns (strong ion exchange resin Q Sepharose XL, weak ion exchange resins, DEAE and ANX, GE Healthcare Biosciences^®, Uppsala, Sweden) pre-equilibrated with 10mM tris-EDTA buffer (pH 8.0). GFP elution range was tested with fractions of a salt gradient from 0.05 to 0.30 M NaCl in 10 mM phosphate buffer (pH 7.0). High concentrations of GFP were eluted in fractions between 0.2 M and 0.3 M NaCl for all resins evaluated. GFP recovery was 26.5%, 62.4% and 139.4% for ANX, DEAE and Q XL matrices, respectively. The Q XL resin resulted in a high loading capacity and better purification, showing that it is well suited for GFP purification, providing a final product with high purity plus high fluorescence intensity.