Multiplexed immunoassay using the stabilized enzymes in mesoporous silica

Yunxian Piao, Dohoon Lee, Jinwoo Lee, Taeghwan Hyeon, Jungbae Kim, Hak Sung Kim

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Multiplexed immunoassay system was developed using the enzyme-immobilized mesoporous silica in a form of nanoscale enzyme reactors (NERs), which improve the enzyme loading, activity, and stability. Glucose oxidase (GO) and trypsin (TR) were adsorbed into mesoporous silica and further crosslinked for the construction of NERs, and antibody-conjugated NERs were employed for the analysis of target antigens in a sandwich-type magnetic bead-based immunoassay. This approach, called as NER-LISA (NER-linked immunosorbent assay), generated signals out of enzyme reactions that correlated well with the concentration of target antigens. The detection limit of NER-LISA using NER-GO and anti-human IgG was 67 pM human IgG, and the sensitivity was 20 times higher than that of the conventional ELISA using anti-human IgG conjugated GO. Antibody-conjugated NER-GO and NER-TR were successfully employed for the simultaneous detection of two target antigens (human IgG and chicken IgG) in a solution by taking advantage of signals at different wavelengths (absorbances at 570 nm and 410 nm, respectively) from the assays of GO and TR activities, demonstrating the potential of NER-LISA in multiplexed immunoassay. The NER-LISA approach also enabled the successful use of a protease (trypsin), because the NER approach can effectively retain the protease molecules within the mesoporous silica and prevent the digestion of antibodies and enzymes during the whole process of NER-LISA.

Original languageEnglish
Pages (from-to)906-912
Number of pages7
JournalBiosensors and Bioelectronics
Volume25
Issue number4
DOIs
Publication statusPublished - 2009 Dec 15

Fingerprint

Immunoassay
Silicon Dioxide
Enzymes
Silica
Glucose Oxidase
Glucose oxidase
Trypsin
Immunoglobulin G
Antigens
Antibodies
Peptide Hydrolases
Assays
Enzyme-Linked Immunosorbent Assay
Immobilized Enzymes
Immunosorbents
Limit of Detection
Digestion
Chickens

Keywords

  • Glucose oxidase
  • Mesoporous silica
  • Multiplexed immunoassay
  • NER-LISA (nanoscale enzyme reactor linked immunosorbent assay)
  • Trypsin

ASJC Scopus subject areas

  • Biophysics
  • Biomedical Engineering
  • Biotechnology
  • Electrochemistry

Cite this

Multiplexed immunoassay using the stabilized enzymes in mesoporous silica. / Piao, Yunxian; Lee, Dohoon; Lee, Jinwoo; Hyeon, Taeghwan; Kim, Jungbae; Kim, Hak Sung.

In: Biosensors and Bioelectronics, Vol. 25, No. 4, 15.12.2009, p. 906-912.

Research output: Contribution to journalArticle

Piao, Yunxian ; Lee, Dohoon ; Lee, Jinwoo ; Hyeon, Taeghwan ; Kim, Jungbae ; Kim, Hak Sung. / Multiplexed immunoassay using the stabilized enzymes in mesoporous silica. In: Biosensors and Bioelectronics. 2009 ; Vol. 25, No. 4. pp. 906-912.
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