|Title:||Aptamer selection and clinical application for C-reactive protein (CRP) based on magnetic-assisted rapid aptamer selection (MARAS)|
Hong Kong Polytechnic University -- Dissertations
|Department:||Faculty of Engineering|
|Pages:||vii, 68 pages : illustrations (some color)|
|Abstract:||Aptamer is one kind of single-strand nucleic acids specifically binding to target of interest and is generally screened by an in vitro method from a random oligonucleotide library using Systematic Evolution of Ligands by EXponential enrichment (SELEX). Utilizing the characteristic of magnetic nanoparticles, Magnetic-Assisted Rapid Aptamer Selection (MARAS) has recently been developed that greatly enhances the efficiency to select aptamers and provides a criteria for selecting aptamers with desired affinity. In this study, specific aptamers for C-reactive protein (CRP) are screened using this straightforward platform, MARAS, with a high affinity of Kd ~ 34 nM. The high specificity of aptamers in this study has been also achieved using multiple runs of negative selection, which is to eliminate the nonspecific binding of aptamers toward non-target biomolecules in serum samples. Selected aptamers are then used as capture ligands for detecting and quantifying the CRP content of forty blind serum samples. These results show a high correlation with those measured using Nephelometry method, one of golden standard to detect CRP using monoclonal anti-CRP antibody in clinical applications. Generally, antibody is necessary for protein detection, however, it is difficult and expensive for a common biological laboratory to produce as the inevitability for an in vivo process. Therefore, this excellent results of the blind test using aptamer-based detection are probably opening a great potential of aptamer to compete with antibody as targets of aptamers are not limited to proteins but antibody does.|
|Rights:||All rights reserved|
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