Generation and Characterization of Murine Monoclonal Antibodies anti-PBP2a of Methicillin-resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a multidrug-resistant bacterium that causes serious infections worldwide. This pathogen is resistant to all beta lactam antibiotics due the presence of PBP2a, a transpeptidase enzyme that presents very low beta-lactam affinity. Here we report the generation and characterization of mouse monoclonal antibodies to PBP2a of MRSA strains. Two clones were obtained and characterized by immunoassays (ELISA, avidity index determination, and immunoblotting), isotyping, association/dissociation rate constants by surface plasmon resonance (SPR), and flow cytometry. Clone 38, which showed the best avidity and affinity, bound to PBP2a located on the bacterial surface by flow cytometry. Further studies are warranted in order to evaluate if these antibodies may help inhibit bacterial growth and be used to treat infections by MRSA.

[1]  A. Tomasz,et al.  Mechanisms of vancomycin resistance in Staphylococcus aureus. , 2014, The Journal of clinical investigation.

[2]  Xuefen Li,et al.  Combined use of the BinaxNOW Staphylococcus aureus test with the Clearview PBP2a assay for the early detection of methicillin-resistant S. aureus from positive blood cultures. , 2014, Diagnostic microbiology and infectious disease.

[3]  G. Corey,et al.  Methicillin-resistant Staphylococcus aureus: an evolving pathogen. , 2014, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[4]  O. Denis,et al.  Evaluation of Clearview Exact PBP2a, a New Immunochromatographic Assay, for Detection of Low-Level Methicillin-Resistant Staphylococcus aureus (LL-MRSA) , 2012, Journal of Clinical Microbiology.

[5]  G. B. Orsi,et al.  Surveillance and management of multidrug-resistant microorganisms , 2011, Expert review of anti-infective therapy.

[6]  S. Ōmura,et al.  Development of an Immunochromatographic Strip for Simple Detection of Penicillin-Binding Protein 2′ , 2010, Clinical and Vaccine Immunology.

[7]  J. Wehland,et al.  Functional Antibodies Targeting IsaA of Staphylococcus aureus Augment Host Immune Response and Open New Perspectives for Antibacterial Therapy , 2010, Antimicrobial Agents and Chemotherapy.

[8]  Charles Y. Tan,et al.  A fully human monoclonal antibody to Staphylococcus aureus iron regulated surface determinant B (IsdB) with functional activity in vitro and in vivo. , 2010, Human antibodies.

[9]  G. Fischer,et al.  Phase 1/2 Double-Blind, Placebo-Controlled, Dose Escalation, Safety, and Pharmacokinetic Study of Pagibaximab (BSYX-A110), an Antistaphylococcal Monoclonal Antibody for the Prevention of Staphylococcal Bloodstream Infections, in Very-Low-Birth-Weight Neonates , 2009, Antimicrobial Agents and Chemotherapy.

[10]  Brook E. Ragle,et al.  Anti-Alpha-Hemolysin Monoclonal Antibodies Mediate Protection against Staphylococcus aureus Pneumonia , 2009, Infection and Immunity.

[11]  G. Yarranton,et al.  Antibodies for the treatment of bacterial infections: current experience and future prospects. , 2008, Current opinion in biotechnology.

[12]  H. Boucher,et al.  Epidemiology of methicillin-resistant Staphylococcus aureus. , 2008, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[13]  Roberta B Carey,et al.  Invasive methicillin-resistant Staphylococcus aureus infections in the United States. , 2007, JAMA.

[14]  D. Machado,et al.  Evaluation of the humoral immune response in BALB/c mice immunized with a naked DNA vaccine anti-methicillin-resistant Staphylococcus aureus. , 2006, Genetics and molecular research : GMR.

[15]  I. Gould,et al.  The clinical significance of methicillin-resistant Staphylococcus aureus. , 2005, The Journal of hospital infection.

[16]  E. Gorovits,et al.  Characterization of a Humanized Monoclonal Antibody Recognizing Clumping Factor A Expressed by Staphylococcus aureus , 2005, Infection and Immunity.

[17]  D. Santos,et al.  Protective immune response against methicillin resistant Staphylococcus aureus in a murine model using a DNA vaccine approach. , 2003, Vaccine.

[18]  Daniel Lim,et al.  Structural basis for the β lactam resistance of PBP2a from methicillin-resistant Staphylococcus aureus , 2002, Nature Structural Biology.

[19]  G. Archer,et al.  Conversion of Oxacillin-Resistant Staphylococci from Heterotypic to Homotypic Resistance Expression , 2002, Antimicrobial Agents and Chemotherapy.

[20]  Daniel Lim,et al.  Structural basis for the beta lactam resistance of PBP2a from methicillin-resistant Staphylococcus aureus. , 2002, Nature structural biology.

[21]  I. Nagaoka,et al.  DNA vaccination by mecA sequence evokes an antibacterial immune response against methicillin-resistant Staphylococcus aureus. , 1999, Journal of Antimicrobial Chemotherapy.

[22]  D. Myszka,et al.  Improving biosensor analysis , 1999, Journal of molecular recognition : JMR.

[23]  J. Ghuysen,et al.  Multimodular Penicillin-Binding Proteins: An Enigmatic Family of Orthologs and Paralogs , 1998, Microbiology and Molecular Biology Reviews.

[24]  A. Tomasz,et al.  Geographic spread of epidemic multiresistant Staphylococcus aureus clone in Brazil , 1995, Journal of clinical microbiology.

[25]  B. Berger-Bächi,et al.  Mechanisms of heteroresistance in methicillin-resistant Staphylococcus aureus , 1994, Antimicrobial Agents and Chemotherapy.