Antibiotic resistance is a global health issue, leading to increased death rates and health costs. In Staphylococcus aureus, resistance to beta-lactams is particularly worrying. Methicillin-resistant S. aureus (MRSA) strains have acquired an additional penicillin-binding protein (PBP), PBP2a, able to synthesize cell wall in the presence of beta-lactams. To treat MRSA infections, ceftaroline was introduced to the market in 2010. A report by Long et al. (2014) found strains highly resistant to ceftaroline in a patient (> 32 mg/L). In this study, we cloned and purified one of the PBP2a mutants conferring ceftaroline resistance, PBP2a-5007. We showed that : i) PBP2a-5007 has a lower affinity to ceftaroline when compared to reference PBP2a-COL ; ii) PBP2a-5007 showed a similar thermal unfolding profile as PBP2a- COL ; iii) PBP2a-5007 showed no resistance to ceftaroline when cloned and expressed on a plasmid in a S. aureus COL ∆mecA strain. Qualification of clavulanate/penicillin G synergy effect on unfolding was attempted. In conclusion, we have produced recombinant PBP2a- 5007 and successfully determined key properties for ceftaroline resistance and stability.