51. Detection of cyclic di-AMP using a competitive ELISA with a unique pneumococcal cyclic di-AMP binding protein
- Author
-
Yang Zhang, Guangchun Bai, Dennis W. Metzger, and Adam J. Underwood
- Subjects
Microbiology (medical) ,Binding protein ,Reproducibility of Results ,Enzyme-Linked Immunosorbent Assay ,Plasma protein binding ,AMP binding ,Biology ,medicine.disease_cause ,Microbiology ,Molecular biology ,Sensitivity and Specificity ,Article ,Bacterial protein ,Streptococcus pneumoniae ,Biochemistry ,Bacterial Proteins ,Carrier protein ,medicine ,Cyclic AMP ,Carrier Proteins ,Molecular Biology ,Protein Binding - Abstract
Cyclic di-AMP (c-di-AMP) is a signaling molecule that has been shown to play important roles in bacterial physiology and infections. Currently, c-di-AMP detection and quantification relies mostly on the use of high-performance liquid chromatography (HPLC) or liquid chromatography-mass spectrometry (LC-MS). In this study, a competitive enzyme-linked immunosorbent assay (ELISA) for the quantification of c-di-AMP was developed, which utilizes a novel pneumococcal c-di-AMP binding protein (CabP) and a newly commercialized c-di-AMP derivative. With this new method, c-di-AMP concentrations in biological samples can be quickly and accurately quantified. Furthermore, this assay is much more efficient than current methods as it requires less overall cost and training while processing many samples at once. Therefore, this assay can be extensively used in research into c-di-AMP signaling.
- Published
- 2014