Miniaturized amperometric flow immunoassay system using a glass fiber membrane modified with anion

Tae Kyu Lim, Satoko Imai, Tadashi Matsunaga

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

This paper describes a miniaturized amperometric flow immunoassay system using a glass fiber membrane modified with anion. The glass fiber membrane was functionally modified with γ-glycidoxypropyltrimethoxysilane and sodium thiosulfate and was used for separation of protein. Anti-human chorionic gonadotrophin (HCG) immunoglobulin G (IgG) antibody conjugated with ferrocenemonocarboxylic acid (Fc), namely, Fc-conjugated IgG (Fc-IgG), was used as a novel analytical reagent. HCG and Fc-IgG complexes were separated from free Fc-IgG based on differences in isoelectric point (pl) using the glass fiber membrane modified with a thiosulfonyl acid functional group. The assay yields a linear relationship between current and HCG concentration in the range of 0-2000 mlU/mL. This simple technique enables the assay of HCG within 2 min. The modified glass fiber membrane was regenerated by occasional elution with malonate buffer (pH 6.0) containing 0.5 M NaCl, to remove free Fc-IgG. Free Fc-IgG recovered in this manner could be reused up to eight times without significant decreases in sensitivity. This miniaturized amperometric flow immunoassay requires only minute quantities of serum and generates highly reproducible results.

Original languageEnglish
Pages (from-to)758-763
Number of pages6
JournalBiotechnology and Bioengineering
Volume77
Issue number7
DOIs
Publication statusPublished - 2002 Mar 30
Externally publishedYes

Keywords

  • Ferrocene-conjugated IgG
  • Glass fiber membrane
  • Human chorionic gonadotrophin
  • Miniaturized amperometric flow immunoassay

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

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