Biopharmaceuticals are gaining high relevance in the pharmaceutical industry and market, due to their specific design for many therapeutic conditions and ability to restore biological functions. As a matter of fact, gene therapy products have been studied and developed in the past several years, and more recently, they arouse real interest as promising alternatives for the establishment of rapid, efficient, and safe treatments for incurable diseases and pandemics. Plasmid DNA (pDNA) is a nonviral vector with great potential in gene therapy due to its biosafety, standard manufacturing, and reduced production cost. Among the different pDNA topologies, the supercoiled (sc) isoform is considered a structure with a higher therapeutic effect. Thus, the production and purification of such a biomolecule must be controlled by suitable analytical methods in order to guarantee the fulfillment of the criteria established by regulatory agencies regarding the purity and homogeneity of sc pDNA for therapeutic applications. Therefore, it is important to ensure that the quality control methods are fast, precise, and specific for the assessment of the target biomolecule. Thus, in the laboratory experiment of a pharmaceutical biotechnology class, a commercial CIMac DEAE pDNA analytical column is used by students for the assessment of the purity level and recovery yield of sc pDNA obtained by recombinant production and a histamine monolith-based purification process. This laboratory class allows students of the Biotechnology or Pharmaceutical Sciences Master degrees to understand the global biotechnological process involved in sc pDNA production and the quality control methods necessary to validate these products for safe therapeutic administration.