Detection and Removal of Escherichia coli Using Fluorescein Isothiocyanate Conjugated Monoclonal Antibody Immobilized on Bacterial Magnetic Particles

Noriyuki Nakamura; J. Grant Burgess; Kaoru Yagiuda; Satoko Kudo; Toshifumi Sakaguchi; Tadashi Matsunaga

doi:10.1021/ac00063a018

Detection and Removal of Escherichia coli Using Fluorescein Isothiocyanate Conjugated Monoclonal Antibody Immobilized on Bacterial Magnetic Particles

Noriyuki Nakamura, J. Grant Burgess, Kaoru Yagiuda, Satoko Kudo, Toshifumi Sakaguchi, Tadashi Matsunaga

Research output: Contribution to journal › Article › peer-review

94 Citations (Scopus)

Abstract

A novel fluoroimmunoassay method using bacterial magnetic particles for the highly sensitive detection of bacteria has been developed. Fluorescein isothiocyanate (FITC) conjugated monoclonal anti-Escherichia coli antibody was immobilized onto bacterial magnetic particles (BMPs) using a heterobifunctional reagent, N-succinimdyl 3-(2-pyridyldithio)propionate (SPDP). E. coli cells were reacted with FITC-antibody-BMP conjugates for 15 min in an inhomogeneous magnetic field which enhanced aggregation. The cell/BMP complexes sedimented, causing relative fluorescence intensity of the solution to decrease with increasing microbial cell concentration. A linear relationship was obtained between the relative fluorescence intensity and cell concentration in the range of 10₂-10₆ cells/mL. Selectivity of this detection system was satisfactory. Monoclonal antibody immobilized on BMPs was also applied to the specific removal of E, coli from the bacterial suspension.

Original language	English
Pages (from-to)	2036-2039
Number of pages	4
Journal	Analytical Chemistry
Volume	65
Issue number	15
DOIs	https://doi.org/10.1021/ac00063a018
Publication status	Published - 1993 Aug 1
Externally published	Yes

ASJC Scopus subject areas

Analytical Chemistry

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10.1021/ac00063a018

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title = "Detection and Removal of Escherichia coli Using Fluorescein Isothiocyanate Conjugated Monoclonal Antibody Immobilized on Bacterial Magnetic Particles",

abstract = "A novel fluoroimmunoassay method using bacterial magnetic particles for the highly sensitive detection of bacteria has been developed. Fluorescein isothiocyanate (FITC) conjugated monoclonal anti-Escherichia coli antibody was immobilized onto bacterial magnetic particles (BMPs) using a heterobifunctional reagent, N-succinimdyl 3-(2-pyridyldithio)propionate (SPDP). E. coli cells were reacted with FITC-antibody-BMP conjugates for 15 min in an inhomogeneous magnetic field which enhanced aggregation. The cell/BMP complexes sedimented, causing relative fluorescence intensity of the solution to decrease with increasing microbial cell concentration. A linear relationship was obtained between the relative fluorescence intensity and cell concentration in the range of 102-106 cells/mL. Selectivity of this detection system was satisfactory. Monoclonal antibody immobilized on BMPs was also applied to the specific removal of E, coli from the bacterial suspension.",

author = "Noriyuki Nakamura and Burgess, {J. Grant} and Kaoru Yagiuda and Satoko Kudo and Toshifumi Sakaguchi and Tadashi Matsunaga",

year = "1993",

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AU - Nakamura, Noriyuki

AU - Burgess, J. Grant

AU - Yagiuda, Kaoru

AU - Kudo, Satoko

AU - Sakaguchi, Toshifumi

AU - Matsunaga, Tadashi

PY - 1993/8/1

Y1 - 1993/8/1

N2 - A novel fluoroimmunoassay method using bacterial magnetic particles for the highly sensitive detection of bacteria has been developed. Fluorescein isothiocyanate (FITC) conjugated monoclonal anti-Escherichia coli antibody was immobilized onto bacterial magnetic particles (BMPs) using a heterobifunctional reagent, N-succinimdyl 3-(2-pyridyldithio)propionate (SPDP). E. coli cells were reacted with FITC-antibody-BMP conjugates for 15 min in an inhomogeneous magnetic field which enhanced aggregation. The cell/BMP complexes sedimented, causing relative fluorescence intensity of the solution to decrease with increasing microbial cell concentration. A linear relationship was obtained between the relative fluorescence intensity and cell concentration in the range of 102-106 cells/mL. Selectivity of this detection system was satisfactory. Monoclonal antibody immobilized on BMPs was also applied to the specific removal of E, coli from the bacterial suspension.

AB - A novel fluoroimmunoassay method using bacterial magnetic particles for the highly sensitive detection of bacteria has been developed. Fluorescein isothiocyanate (FITC) conjugated monoclonal anti-Escherichia coli antibody was immobilized onto bacterial magnetic particles (BMPs) using a heterobifunctional reagent, N-succinimdyl 3-(2-pyridyldithio)propionate (SPDP). E. coli cells were reacted with FITC-antibody-BMP conjugates for 15 min in an inhomogeneous magnetic field which enhanced aggregation. The cell/BMP complexes sedimented, causing relative fluorescence intensity of the solution to decrease with increasing microbial cell concentration. A linear relationship was obtained between the relative fluorescence intensity and cell concentration in the range of 102-106 cells/mL. Selectivity of this detection system was satisfactory. Monoclonal antibody immobilized on BMPs was also applied to the specific removal of E, coli from the bacterial suspension.

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Detection and Removal of Escherichia coli Using Fluorescein Isothiocyanate Conjugated Monoclonal Antibody Immobilized on Bacterial Magnetic Particles

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