An antibody binding-based fluorescent assay for the rapid quantification of globotriaosylceramide levels in human Fabry cells

Allan Wee Ren Ng; Kumaran Narayanan

doi:10.1016/j.ab.2021.114287

An antibody binding-based fluorescent assay for the rapid quantification of globotriaosylceramide levels in human Fabry cells

Allan Wee Ren Ng
, Kumaran Narayanan

Malaysia Sch of Science

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

1 Citation (Scopus)

Abstract

Fabry disease is caused by reduced α-GAL A activity and accumulation of globotriaosylceramide (Gb₃). Here, we describe a microplate Gb₃ assay using fluorophore-tagged antibody and crude cellular lipid extracts. The assay is able to detect higher Gb₃ concentrations in human Fabry cells compared to non-diseased cells. This result was verified by immunofluorescence staining that revealed large amounts of Gb₃ deposits in Fabry cell lines, demonstrating the accuracy of this method. This assay may provide the basis for detecting Fabry disease by quantifying Gb₃ deposits from human biological samples, for example, from urine and blood.

Original language	English
Article number	114287
Number of pages	4
Journal	Analytical Biochemistry
Volume	628
DOIs	https://doi.org/10.1016/j.ab.2021.114287
Publication status	Published - 1 Sept 2021

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Diagnosis
Fabry disease
Globotriaosylceramide
Quantification
α-galactosidase A

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10.1016/j.ab.2021.114287

Cite this

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title = "An antibody binding-based fluorescent assay for the rapid quantification of globotriaosylceramide levels in human Fabry cells",

abstract = "Fabry disease is caused by reduced α-GAL A activity and accumulation of globotriaosylceramide (Gb3). Here, we describe a microplate Gb3 assay using fluorophore-tagged antibody and crude cellular lipid extracts. The assay is able to detect higher Gb3 concentrations in human Fabry cells compared to non-diseased cells. This result was verified by immunofluorescence staining that revealed large amounts of Gb3 deposits in Fabry cell lines, demonstrating the accuracy of this method. This assay may provide the basis for detecting Fabry disease by quantifying Gb3 deposits from human biological samples, for example, from urine and blood.",

keywords = "Diagnosis, Fabry disease, Globotriaosylceramide, Quantification, α-galactosidase A",

author = "Ng, \{Allan Wee Ren\} and Kumaran Narayanan",

note = "Funding Information: This study was supported by a Fundamental Research Grant Scheme FRGS/1/2019/STG05/MUSM/02/2 provided by the Ministry of Higher Education, Malaysia to K.N. A.W.R.N is supported by Monash Postgraduate Merit Scholarship offered by Monash University Malaysia . Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",

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doi = "10.1016/j.ab.2021.114287",

language = "English",

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T1 - An antibody binding-based fluorescent assay for the rapid quantification of globotriaosylceramide levels in human Fabry cells

AU - Ng, Allan Wee Ren

AU - Narayanan, Kumaran

N1 - Funding Information: This study was supported by a Fundamental Research Grant Scheme FRGS/1/2019/STG05/MUSM/02/2 provided by the Ministry of Higher Education, Malaysia to K.N. A.W.R.N is supported by Monash Postgraduate Merit Scholarship offered by Monash University Malaysia . Publisher Copyright: © 2021 Elsevier Inc.

PY - 2021/9/1

Y1 - 2021/9/1

N2 - Fabry disease is caused by reduced α-GAL A activity and accumulation of globotriaosylceramide (Gb3). Here, we describe a microplate Gb3 assay using fluorophore-tagged antibody and crude cellular lipid extracts. The assay is able to detect higher Gb3 concentrations in human Fabry cells compared to non-diseased cells. This result was verified by immunofluorescence staining that revealed large amounts of Gb3 deposits in Fabry cell lines, demonstrating the accuracy of this method. This assay may provide the basis for detecting Fabry disease by quantifying Gb3 deposits from human biological samples, for example, from urine and blood.

AB - Fabry disease is caused by reduced α-GAL A activity and accumulation of globotriaosylceramide (Gb3). Here, we describe a microplate Gb3 assay using fluorophore-tagged antibody and crude cellular lipid extracts. The assay is able to detect higher Gb3 concentrations in human Fabry cells compared to non-diseased cells. This result was verified by immunofluorescence staining that revealed large amounts of Gb3 deposits in Fabry cell lines, demonstrating the accuracy of this method. This assay may provide the basis for detecting Fabry disease by quantifying Gb3 deposits from human biological samples, for example, from urine and blood.

KW - Diagnosis

KW - Fabry disease

KW - Globotriaosylceramide

KW - Quantification

KW - α-galactosidase A

UR - https://www.scopus.com/pages/publications/85107953093

U2 - 10.1016/j.ab.2021.114287

DO - 10.1016/j.ab.2021.114287

M3 - Article

C2 - 34119486

AN - SCOPUS:85107953093

SN - 0003-2697

VL - 628

JO - Analytical Biochemistry

JF - Analytical Biochemistry

M1 - 114287

ER -

An antibody binding-based fluorescent assay for the rapid quantification of globotriaosylceramide levels in human Fabry cells

Abstract

UN SDGs

Keywords

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