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Background and Objectives Discovery of antibiotics has been one of the important achievements of mankind in the field of infectious diseases, although after a short time, the phenomenon of antibiotic resistance was created. Nowadays, there is a need for an innovation in the control of microbial infections such as the use of antimicrobial peptides. These peptides are abundantly found in neutrophils and since leukoreduction filters are a valuable source of these cells, it has been studied as an available option Materials and Methods In this experimental study, leukoreduction filters was selected by easy non-probability sampling method. First, leukocytes were extracted from leukoreduction filters and then neutrophils were isolated. Defensin peptide was extracted after sonication of neutrophils and was purified by HPLC. Staphylococcus aureus bacteria was cultured and the amount of bacterial inoculation was determined and the minimum inhibitory concentration was measured by macrodilution method. Microbial inhibition halo diameter size of defensin was determined for the intended bacteria. Results Using defensin standard, purified defensin peak was collected and the minimum inhibitory concentration was determined as 2.5 μg/mL on Staphylococcus aureus by macrodilution method. Microbial inhibition halo diameter size was calculated as 9 mm Conclusions The treatment of infectious diseases has faced a challenge with the widespread emergence of drug resistance, and the use of new innovations is inevitable. Defensin as an antimicrobial peptide is a suitable option that is abundantly found in neutrophils obtained from leukoreduction filters and is suggested for research on new antimicrobial drugs. [ABSTRACT FROM AUTHOR]