원문정보
The Method Development for Biomarker Diagnosis Based on the Aptamer-protein Crosslink
초록
영어
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.
목차
1. 서론
2. 재료 및 방법
2.1. 실험 재료
2.2. 수정된 면역침강법을 이용한 목적 단백질 검출
2.3. 질량분석기를 이용한 단백질 분석
3. 결과 및 고찰
3.1. 앱타머와 마그네틱 비드를 이용한 수정된 크로마틴 면역침강법 응용
3.2. 목적 단백질 검출을 위한 반응 최적화
3.3. 질병 진단에의 응용을 위한 검출 한도 확인
4. 결론
감사
References