Immuno-capture of Cryptosporidium parvum using micro-well array

Tomoyuki Taguchi, Haruko Takeyama, Tadashi Matsunaga

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

A glass slide and micro-well array chip on which anti-Cryptosporidium parvum antibody was immobilized were used for the rapid capture and detection of C. parvum. Biotinylated anti-C. parvum antibodies were spotted onto the streptavidin-coated glass slides. C. parvum oocysts were captured specifically on the spot when more than 73 ng of anti-C. parvum antibody was applied onto the glass slide. However, C. parvum oocysts captured on the glass slide were detached by repeating washing steps. To improve the capture efficiency of oocysts, capture was performed in a micro-well format consisting of 1024 wells/2.5 mm2 (32 × 32 wells) fabricated as a chip by photolithography. Instead of a flat surface on a glass slide, each well was 30 μm in diameter and 10 μm in depth. Streptavidin was also immobilized onto the micro-well array. The biotinylated anti-C. parvum antibodies were immobilized efficiently onto the chip using a buffer containing 20% methanol. Using this technique C. parvum oocysts were stably captured onto the chip after repeated washing procedures. These data show that the newly designed micro-well array technique described here is useful for antibody-mediated C. parvum capture.

Original languageEnglish
Pages (from-to)2276-2282
Number of pages7
JournalBiosensors and Bioelectronics
Volume20
Issue number11
DOIs
Publication statusPublished - 2005 May 15
Externally publishedYes

Keywords

  • Cryptosporidium parvum
  • Immuno-capture
  • Micro-well array
  • Microfabrication

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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