The development of sandwich-type binding pair of aptamers-based electrochemical and lateral flow biosensors
샌드위치형 결합 쌍앱타머 기반의 전기화학 및 측방-유동 바이오센서 개발 연구
- 주제(키워드) Aptamer , Biosensor
- 발행기관 고려대학교 대학원
- 지도교수 구만복
- 발행년도 2020
- 학위수여년월 2020. 2
- 학위구분 박사
- 학과 대학원 생명공학과
- 세부전공 의약나노바이오시스템공학
- 원문페이지 113 p
- UCI I804:11009-000000127143
- DOI 10.23186/korea.000000127143.11009.0000950
- 본문언어 영어
- 제출원본 000046025490
초록/요약
Since the first development of biosensor in 1962, the field of biosensor research has grown remarkably with convergence of micro/nanotechnology, electronics, material science and basic sciences. In order to develop more sensitive, selective, and robust biosensor, choosing appropriate recognition element, assay-types, and platforms are important. Aptamer is a notable biorecognition element and widely used for biosensor due to its various advantages features over other receptors. Most commonly used assay-type is sandwich-type assay since it is more sensitive and selective compared to the competitive-type assay. Moreover, when using sandwich-type assay, signal amplification is eligible by modifying the secondary bioreceptor with enzymes or nanomaterials. Considering the biosensing transducer, electrochemical-based and lateral flow-based platforms are noteworthy since those are already used for commercialized point-of-care diagnostics. In this research, the development of sandwich-type electrochemical and lateral flow biosensors using pair of aptamers for type 2 diabetes biomarker and avian influenza virus were discussed. In case of type 2 diabetes biomarker detection, aptamer duo was applied to the screen-printed gold electrode (SPGE). Further, to improve the sensitivity, surface modified gold electrode with coccolith was investigated. The newly developed coccolith modified electrode-based aptasensor showed increased signal intensity compared to the bare SPGE. Moreover, modified electrode showed the 3-fold enhanced limit of detection (LOD) in serum condition. For avian influenza virus detection, avian influenza virus pair of aptamers were applied to lateral flow assay for the specific detection of the virus. This sandwich-type aptamer-based lateral flow biosensor showed LOD of 6x105 EID50/ml and 1.2x106 EID50/ml in buffer and in duck’s feces condition, respectively, which are similar to the commercially available rapid kit. Regarding the advantages of sandwich-type format and aptamer, these studies could pave the way for the development of noble biosensors for point-of-care diagnostics.
more초록/요약
바이오센서가 처음 개발된 이후로 바이오센서분야는 나노과학, 재료과학 및 전자과학과 융합되면서 꾸준히 성장하였다. 바이오센서를 구성하는 요소는 크게 바이오인식물질과 신호변환플랫폼이 있지만, 이외에도 검출방식 또한 매우 중요하다. 따라서, 검출 성능이 뛰어나고 안정성이 높은 바이오센서 개발을 위해서는 바이오센서의 중요 요소인 바이오인식물질, 신호변환플랫폼 및 검출방식을 적절히 고려하는 것이 중요하다. 여러 바이오인식물질 중 앱타머는 최근 여러 바이오센서의 개발에서 널리 이용되고 있으며, 그 자체의 특성으로 인해 다른 바이오리셉터와 비교하여 뛰어난 장점을 가지고 있기에 주목할 만한 바이오인식 물질이다. 검출방식으로는 샌드위치 형태의 방식이 주로 이용되는데, 이는 샌드위치 방식이 경합측정방식과 비교하였을 때, 민감도와 선택성 측면에서 더욱 좋기 때문이다. 또한, 샌드위치 방식을 사용하게 되면, 효소 및 나노입자를 이용하여 신호증폭이 용이하기에 검출하고자 하는 타겟을 더욱 민감하게 측정할 수 있다. 바이오센서 플랫폼 측면에서는 현장진단측면에서 가장 널리 이용되는 전기화학적 플랫폼과 측방유동 플랫폼이 주목할 만하다.
more목차
1. Chapter - Introduction 1
1.1. Overview 2
1.2. Literature survey 3
1.2.1. ssDNA Aptamer 3
1.2.2. Aptamer-based Optical biosensing platforms 6
1.2.3. Aptamer-based electrochemical biosensor 10
1.2.4. Other types of aptamer-based biosensor platforms 12
2. Chapter Cognate pair of aptamer-based sandwich-type electrochemical biosensor for vaspin 14
2.1. Introduction 15
2.2. Materials and Methods 16
2.2.1. Materials 16
2.2.2. Apparatus and electrochemical measurements 17
2.2.3. Preparation of a sandwich-type aptamer-based biosensor for vaspin 17
2.2.4. Assay procedure for vaspin detection using a sandwich-type aptamer-based biosensor 18
2.3. Results and Discussion 19
2.3.1. Characteristics of sandwich-type aptamer-based electrochemical biosensor 20
2.3.2. Analytical performance of the sandwich-type aptamer-based electrochemical biosensor 22
2.4. Conclusion 25
3. Chapter Coccolith shell modified electrode-based sandwich-type electrochemical biosensor for vaspin 26
3.1. Introduction 27
3.2. Materials and Methods 29
3.2.1. Materials 29
3.2.2. Apparatus and electrochemical measurements 30
3.2.3. Preparation of Coccolith Shells 31
3.2.4. Preparation of Coccolith modified electrodeposited SPGE (CME-SPGE) 32
3.2.5. Preparation of the sandwich-type electrochemical biosensor using a cognate pair of aptamers and assay procedure 34
3.2.6. Characterization 36
3.3. Results and Discussion 37
3.3.1. Morphological Characterization of the Coccolith modified SPGE 38
3.3.2. Electrochemical properties of the Coccolith modified electrodeposited SPGE 41
3.3.3. Characteristics of the Successfully developed Sandwich-type Electrochemical Aptasensor 44
3.3.4. Analytical Performance of the Sandwich-type Electrochemical Biosensor using a Cognate Pair of Aptamers 46
3.4. Conclusion 52
4. Chapter - Cognate pair of aptamer-based sandwich-type lateral flow biosensor for avian influenza virus Avian influenza virus detection using cognate pair of aptamers 53
4.1. Introduction 54
4.2. Materials and Methods 58
4.2.1. Materials 58
4.2.2. Cognate pair of aptamer-based lateral flow strip biosensor 59
4.3. Results and Discussion 60
Development of a sandwich-type lateral flow strip biosensor using a cognate pair of aptamers for the whole virus 62
4.4. Conclusion 69
5. Chapter – Concluding Remarks 71
5.1. Summary and Conclusion 71
5.2. Further study 73
6. References 74
국문 초록 98
Acknowledgement 100
