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KMID : 0380020110260040352
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Korean Journal of Biotechnology and Bioengineering
2011 Volume.26 No. 4 p.352 ~ p.356
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The Method Development for Biomarker Diagnosis Based on the Aptamer-protein Crosslink
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Lee Bo-Rahm
Kim Ji-Nu
Kim Byung-Gee
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Abstract
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The detection of biomarkers is an important issue for disease diagnosis. However, many systems are not suitable to detect the biomarker itself directly. For direct detection of biomarker proteins in human serum, a new affinity-capture method using aptamers combined with the mass spectrometry was suggested. Since signals from protein samples cannot be amplified, modified chromatin immunoprecipitation (ChIP) and subsequent cross-linking with formaldehyde between aptamers and target proteins were used not to lose the captured target proteins, which allowed us to perform a harsh washing step to remove the non-specifically bound proteins. As a model system, a thrombin aptamer was used as a bait and thrombin as a target protein. Using our modified ChIP and affinity-capture method, non-specific binding proteins on the beads decreased significantly, suggesting that our new method is efficient and can be applied to developing diagnosis systems for various biomarkers.
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KEYWORD
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aptamer biomarker, formaldehyde, cross-linking, MS analysis, magnetic beads
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