앱타머와 단백질간 가교를 이용한 바이오마커 진단 방법 개발

• Published in: KSBB Journal Vol. 26; no. 4; pp. 352 - 356
• Main Authors: 이보람, 김진우, 김병기
• Format: Journal Article
• Language: Korean
• Published: 한국생물공학회 2011
• Subjects: 생물공학; MS analysis; formaldehyde; magnetic beads; cross-linking; aptamer biomarker
• ISSN: 1225-7117; 2288-8268

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.