Abstract
In this protocol, we describe the application of using a high affinity monoclonal antibody generated against the major pilin protein component of the pilin structure of Enterococcus faecalis as a PET imaging agent for enterococcal endocarditis detection. The anti-pilin -mAb 64Cu conjugate was able to specifically label enterococcal endocarditis vegetation in vivo in a rodent endocarditis model. By targeting pili, a covalently linked surface antigen extending from the bacterial surface, we provided evidence that gram-positive pilin represent a logical surface antigen to define or target an infectious agent for molecularly guided imaging. Our goal in providing a detailed protocol of our efforts is to enable others to build upon this methodology to answer pertinent translational and basic research questions in the pursuit of diagnosis and treatment of infective endocarditis.
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References
Lerche CJ et al (2015) Low efficacy of tobramycin in experimental Staphylococcus aureus endocarditis. Eur J Clin Microbiol Infect Dis 34(12):2349–2357
Hanses F et al (2014) Staphylococcus aureus gene expression in a rat model of infective endocarditis. Genome Med 6(10):93
Jung CJ et al (2009) Streptococcus mutans autolysin AtlA is a fibronectin-binding protein and contributes to bacterial survival in the bloodstream and virulence for infective endocarditis. Mol Microbiol 74(4):888–902
Entenza JM et al (2005) Therapeutic effects of bacteriophage Cpl-1 lysin against Streptococcus pneumoniae endocarditis in rats. Antimicrob Agents Chemother 49(11):4789–4792
Strohl WR (2009) Optimization of Fc-mediated effector functions of monoclonal antibodies. Curr Opin Biotechnol 20(6):685–691
Cao Y et al (2012) Single-chain antibody-based immunotoxins targeting Her2/neu: design optimization and impact of affinity on antitumor efficacy and off-target toxicity. Mol Cancer Ther 11(1):143–153
Hall MA et al (2012) Comparison of mAbs targeting epithelial cell adhesion molecule for the detection of prostate cancer lymph node metastases with multimodal contrast agents: quantitative small-animal PET/CT and NIRF. J Nucl Med 53(9):1427–1437
Gao P et al (2015) Deglycosylation of mAb by EndoS for improved molecular imaging. Mol Imaging Biol 17(2):195–203
Gao P et al (2010) Enterococcus faecalis rnjB is required for pilin gene expression and biofilm formation. J Bacteriol 192(20):5489–5498
Pinkston KL et al (2011) The Fsr quorum-sensing system of Enterococcus faecalis modulates surface display of the collagen-binding MSCRAMM Ace through regulation of gelE. J Bacteriol 193(17):4317–4325
Kao D et al (2015) A Monosaccharide residue is sufficient to maintain mouse and human IgG subclass activity and directs IgG effector functions to cellular Fc receptors. Cell Rep 13(11):2376–2385
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Pinkston, K.L. et al. (2017). Antibody Guided Molecular Imaging of Infective Endocarditis. In: Nordenfelt, P., Collin, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology, vol 1535. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6673-8_15
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DOI: https://doi.org/10.1007/978-1-4939-6673-8_15
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6671-4
Online ISBN: 978-1-4939-6673-8
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