A rapid sandwich immunoassay for human fetuin A using agarose-3-aminopropyltriethoxysilane modified microtiter plate

Sandeep Kumar Vashist; E. Marion Schneider; John H.T. Luong

doi:10.1016/j.aca.2015.04.060

A rapid sandwich immunoassay for human fetuin A using agarose-3-aminopropyltriethoxysilane modified microtiter plate

Sandeep Kumar Vashist
, E. Marion Schneider
, John H.T. Luong

School of Chemistry

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

Abstract

A rapid sandwich immunoassay (IA) with enhanced signal response for human fetuin A (HFA) was developed by modifying the surface of a KOH-treated polystyrene microtiter plate (MTP) with agarose and 3-aminopropyltriethoxysilane (APTES). The agarose-APTES complex binds covalently to the hydroxyl moiety of the MTP plate to serve as a binding platform for bioconjugation of EDC-activated anti-HFA antibody (Ab) via carbodiimide coupling. The one-step kinetics-based sandwich enzyme-linked immunosorbent assay (ELISA) enabled the detection of HFA in 30min with a limit of detection (LOD) and a linear range of 0.02 ng mL^-1 and 1-243 ng mL^-1, respectively. It detected HFA spiked in diluted human whole blood and serum, and HFA in ethylenediaminetetraacetic acid (EDTA)-plasma of patients with high precision similar to that of conventional ELISA. The anti-HFA Ab-bound agarose-functionalized MTPs retained their functional activity after 6 weeks of storage in 0.1M PBS, pH 7.4 at 4°C.

Original language	English
Pages (from-to)	74-80
Number of pages	7
Journal	Analytica Chimica Acta
Volume	883
DOIs	https://doi.org/10.1016/j.aca.2015.04.060
Publication status	Published - 9 Jul 2015

Keywords

Agarose
Human fetuin A
One-step kinetics
Rapid sandwich immunoassay
Signal enhancement

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10.1016/j.aca.2015.04.060

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title = "A rapid sandwich immunoassay for human fetuin A using agarose-3-aminopropyltriethoxysilane modified microtiter plate",

abstract = "A rapid sandwich immunoassay (IA) with enhanced signal response for human fetuin A (HFA) was developed by modifying the surface of a KOH-treated polystyrene microtiter plate (MTP) with agarose and 3-aminopropyltriethoxysilane (APTES). The agarose-APTES complex binds covalently to the hydroxyl moiety of the MTP plate to serve as a binding platform for bioconjugation of EDC-activated anti-HFA antibody (Ab) via carbodiimide coupling. The one-step kinetics-based sandwich enzyme-linked immunosorbent assay (ELISA) enabled the detection of HFA in 30min with a limit of detection (LOD) and a linear range of 0.02 ng mL-1 and 1-243 ng mL-1, respectively. It detected HFA spiked in diluted human whole blood and serum, and HFA in ethylenediaminetetraacetic acid (EDTA)-plasma of patients with high precision similar to that of conventional ELISA. The anti-HFA Ab-bound agarose-functionalized MTPs retained their functional activity after 6 weeks of storage in 0.1M PBS, pH 7.4 at 4°C.",

keywords = "Agarose, Human fetuin A, One-step kinetics, Rapid sandwich immunoassay, Signal enhancement",

author = "Vashist, \{Sandeep Kumar\} and Schneider, \{E. Marion\} and Luong, \{John H.T.\}",

note = "Publisher Copyright: {\textcopyright} 2015 Elsevier B.V..",

year = "2015",

month = jul,

day = "9",

doi = "10.1016/j.aca.2015.04.060",

language = "English",

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AU - Vashist, Sandeep Kumar

AU - Schneider, E. Marion

AU - Luong, John H.T.

PY - 2015/7/9

Y1 - 2015/7/9

N2 - A rapid sandwich immunoassay (IA) with enhanced signal response for human fetuin A (HFA) was developed by modifying the surface of a KOH-treated polystyrene microtiter plate (MTP) with agarose and 3-aminopropyltriethoxysilane (APTES). The agarose-APTES complex binds covalently to the hydroxyl moiety of the MTP plate to serve as a binding platform for bioconjugation of EDC-activated anti-HFA antibody (Ab) via carbodiimide coupling. The one-step kinetics-based sandwich enzyme-linked immunosorbent assay (ELISA) enabled the detection of HFA in 30min with a limit of detection (LOD) and a linear range of 0.02 ng mL-1 and 1-243 ng mL-1, respectively. It detected HFA spiked in diluted human whole blood and serum, and HFA in ethylenediaminetetraacetic acid (EDTA)-plasma of patients with high precision similar to that of conventional ELISA. The anti-HFA Ab-bound agarose-functionalized MTPs retained their functional activity after 6 weeks of storage in 0.1M PBS, pH 7.4 at 4°C.

AB - A rapid sandwich immunoassay (IA) with enhanced signal response for human fetuin A (HFA) was developed by modifying the surface of a KOH-treated polystyrene microtiter plate (MTP) with agarose and 3-aminopropyltriethoxysilane (APTES). The agarose-APTES complex binds covalently to the hydroxyl moiety of the MTP plate to serve as a binding platform for bioconjugation of EDC-activated anti-HFA antibody (Ab) via carbodiimide coupling. The one-step kinetics-based sandwich enzyme-linked immunosorbent assay (ELISA) enabled the detection of HFA in 30min with a limit of detection (LOD) and a linear range of 0.02 ng mL-1 and 1-243 ng mL-1, respectively. It detected HFA spiked in diluted human whole blood and serum, and HFA in ethylenediaminetetraacetic acid (EDTA)-plasma of patients with high precision similar to that of conventional ELISA. The anti-HFA Ab-bound agarose-functionalized MTPs retained their functional activity after 6 weeks of storage in 0.1M PBS, pH 7.4 at 4°C.

KW - Agarose

KW - Human fetuin A

KW - One-step kinetics

KW - Rapid sandwich immunoassay

KW - Signal enhancement

UR - https://www.scopus.com/pages/publications/84931008443

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DO - 10.1016/j.aca.2015.04.060

M3 - Article

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AN - SCOPUS:84931008443

SN - 0003-2670

VL - 883

SP - 74

EP - 80

JO - Analytica Chimica Acta

JF - Analytica Chimica Acta

ER -

A rapid sandwich immunoassay for human fetuin A using agarose-3-aminopropyltriethoxysilane modified microtiter plate

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Keywords

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