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최신연구동향 I, Chair : Eun Yeol LEE (Kyung Hee Univ., Korea)
Bi-Functional Surface Modification and Collapsible Roof Device for Cell-Secreted Cytokine Immunoassay
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- 발행기관
- 한국생물공학회 바로가기
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- 간행물
- 한국생물공학회 학술대회 바로가기
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- 통권
- 2012 춘계학술대회 및 국제심포지움 (2012.04)바로가기
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- 페이지
- pp.128-128
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- 저자
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- 언어
- 한국어(KOR)
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- URL
- https://www.earticle.net/Article/A174320
※ 원문제공기관과의 협약기간이 종료되어 열람이 제한될 수 있습니다.
원문정보
초록
- 영어
- It is often desirable to sequester cells in specific locations on the surface and to integrate sensing elements next to the cells. In the present study, surfaces were fabricated so as to position cytokine sensing domains inside non-fouling poly(ethylene glycol) (PEG) hydrogel microwells. Our aim was to increase sensitivity of micropatterned cytokine immunoassays through covalent attachment of biorecognition molecules. To achieve this, glass substrates were functionalized with a binary mixture of acrylate- and thiol-terminated methoxysilanes. During subsequent hydrogel photopatterning step acrylate moieties served to anchor hydrogel microwells to glass substrates. Importantly, glass attachment sites within the microwells contained thiol groups that could be activated with a hetero-bifunctional cross-linker for covalent immobilization of proteins. After incubation with fluorescently-labeled avidin, microwells fabricated on a mixed acryl/thiol silane layer emitted ~6 times more fluorescence compared to microwells fabricated on an acryl silane alone. This result highlighted the advantages of covalent attachment of avidin inside the microwells. To create cytokine immunoassays, micropatterned surfaces were incubated with biotinylated IFN-g or TNF-a antibodies (Abs). Micropatterned immunoassays prepared in this manner were sensitive down to 1 ng/ml or 60 pM IFN-g. To further prove utility of this bionterface design, macrophages were seeded into 30 mm diameter microwells fabricated on either bi-functional (acryl/thiol) or monofunctional silane layers. Both types of microwells were coated with avidin and biotin-anti-TNF-a prior to cell seeding. Short mitogenic activation followed by immunostaining for TNF-a revealed that microwells created on bi-functional silane layer had 3 times higher signal due to macrophage-secreted TNF-a compared to microwells fabricated on mono-functional silane. The rational design of cytokine-sensing surfaces described here will be leveraged in the future for rapid detection of multiple cytokines secreted by individual immune cells.
키워드
cytokine detection immunoassay surface micropatterning hydrogel microwells collapsible roof device저자
간행물 정보
발행기관
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- 발행기관명
- 한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
- 설립연도
- 1984
- 분야
- 공학>생물공학
- 소개
- 이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다 1. 생물공학 분야의 발전을 위한 연구 협력 2. 생물공학의 실용화를 촉진시키기 위한 산학 협동 3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최 4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간 5. 생물공학 발전을 위한 정책 건의 6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동
간행물
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- 간행물명
- 한국생물공학회 학술대회
- 간기
- 반년간
- 수록기간
- 1985~2013
- 십진분류
- KDC 476 DDC 576
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