Journal
JOURNAL OF BIOMOLECULAR SCREENING
Volume 16, Issue 2, Pages 266-271Publisher
SAGE PUBLICATIONS INC
DOI: 10.1177/1087057110391787
Keywords
aptamer; protective antigen; ELISA; Bacillus anthracis; anthrax
Funding
- Hanyang University [HYU-2008-5]
- Rural Development Administration, the Republic of Korea [20080401034006]
- Baylor University
- National Research Foundation of Korea [R31-2011-000-10010-0] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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The protective antigen (PA) of Bacillus anthracis is a secreted protein that functions as a critical virulence factor. Protective antigen has been selected as a biomarker in detecting bacterial infection. The in vitro selection method, systematic evolution of ligands by exponential enrichment (SELEX), was used to find single-stranded DNAs that were tightly bound to PA. After 8 rounds of the SELEX process with PA, 4 different oligonucleotides (referred to as aptamers) that contain a 30-residue ssDNA sequence were identified. Dissociation constant (K-d) values with Cy3-attached aptamers were determined via fluorophotometry to be within a nanomolar range. The authors attempted to visualize the detection of PA using an aptamer-based enzyme-linked immunosorbent assay method, which has proven to be successful within a nanomolar K-d value range. Furthermore, 2 of the 4 aptamers exhibited specificity to PA against bovine serum albumin and bovine serum. The results of this study demonstrate the analytical potential of an oligonucleotide-based biosensor for a wide variety of applications, particularly in diagnosing disease through specific protein biomarkers. (Journal of Biomolecular Screening 2011;16:266-271)
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