Development of the cell culture monitoring system using a smartphone-mountable paper-based analytical device
- 주제(키워드) Biosensor
- 발행기관 아주대학교
- 지도교수 윤현철
- 발행년도 2017
- 학위수여년월 2017. 2
- 학위명 석사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000024262
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
We developed a biosensing system for the facile monitoring of animal cell culture using paper-based analytical device (PAD) and a smartphone. Because that the animal cells consume glucose and produce lactate under the anaerobic culturing condition, the PAD was designed to simultaneously quantify the concentrations of glucose and lactate in culture media for cell culture monitoring. As a biosensing strategy, the enzyme-mediated colorimetric assay method, utilizing oxidase, horseradish peroxidase (HRP) and chromogenic substrate, were employed. On the PAD, two types of enzyme pairs including glucose oxidase (GOx)-HRP pair and lactate oxidase (LOx)-HRP pair were immobilized with colorless chromophores. In this condition, by the cascadic enzyme reactions among target substrates, oxidases, HRP and chromophore, a concentration-dependent colorimetric (blue) signals is developed on the PAD. With a smartphone gadget designed to aid the image acquisition, images of the PAD were acquired using a mobile application, and the color intensities were quantified as sensor signals. For the glucose assay, the developed biosensing system exhibited 0.3 mM of detection limit (LOD) and 0.9 mM of quantification limit (LOQ) in the dynamic detection range from 0.3 to 8.0 mM of glucose. For lactate biosensing, the LOD and LOQ values were respectively registered as 0.02 mM and 0.06 mM in the dynamic detection range from 0.02 to 0.5 mM of lactate. With the developed biosensing system, simultaneous quantification of glucose and lactate in cell culture media were conducted, exhibiting highly accurate and reproducible results. Based on the results, we propose that the developed optical sensing system using paper-based fluidics is feasible for practical monitoring of animal cell culture.
more목차
1. Introduction
1.1 Point-of-care testing…………………………………………1
1.2 Animal cell culture monitoring system………………………2
1.3 Paper-based analytical device………………………………4
1.4 Utilization of a smartphone as a signal transducer………11
1.5 Aim of thesis……………………………………………………15
2. Materials and methods
2.1 Materials and apparatus………………………………………16
2.2 Fabrication of PAD for glucose and lactate analysis
2.2.1 Formation of fluidic channels on paper…………………16
2.2.2 Immobilization of enzymes and chromophores………17
2.2.3 Attachment of layers with cellulose powder…………18
2.3 Manipulation of the smartphone gadget……………………18
2.4 Glucose and lactate analysis employing enzymatic colorimetric assays
2.4.1 Assay of buffer spiked analytes…………………………19
2.4.2 Assay for cell culture media………………………………19
3. Results and discussion
3.1 Optimization of enzymatic reaction time…………………20
3.2 Quantitative assay
3.2.1 Difference of color intensity depending on concentration………………………………………………………24
3.2.2 Cross-reactivity of oxidases……………………………26
3.2.3 Calibration curves of each analyte………………………26
3.3 Glucose and lactate assays using cell culture media……31
4. Conclusions………………………………………………………40
5. References…………………………………………………………41
