We studied the capacity of an API-rHBsAg purification process to eliminate DNA contamination from yeast-host cell. Firstly, was demonstrated consistency of manufacturing purification process to remove DNA, from (3.9 ± 1.9)10⁸ pg/dose in starting material to (3.4 ± 1.6) pg/dose, equivalent to 8.2 log in Active Pharmaceutical Ingredient (API), measuring DNA quantity in several unit operations along manufacturing process for twenty batches, five consecutive in 2000, 2001, 2003 and 2005. These values for API, lower than 10 pg/dose, accomplish current WHO requirements for Hepatitis B vaccines obtaining by recombinant DNA technology (WHO, 1989; European Pharmacopoeia, 2001a). The main removal factor for manufacturing process, equivalent to 6.4-log, was reached in negative anion-exchange chromatography. Then, the capacity of immunoaffinity chromatography and positive anion-exchange chromatography to remove chromosomal DNA purified from yeast-host cell was assessed using a scaled-down chromatographic process which was shown to yield product meeting purity criteria set for the manufacturing process. Log₁₀ reductions for DNA through the immunoaffinity chromatography and positive anion-exchange chromatography were 7.3 ± 0.1, and 5.8 ± 0.1 respectively. Overall, these studies indicate that total DNA clearance factor for API-rHBsAg manufacturing process was 19.4 log, 2.4 times higher than the real DNA contamination, indicating that API-rHBsAg manufacturing as described here have sufficient DNA reducing capacity to achieved a high margin of DNA safety.