Feasibility Study of Anti-Neurofilament Antibodies Detection by Indirect Quenching Fluoroimmunoassay

Alireza  Nakhaee; Manuchehr Messripour; Alireza Khosravi; Saeedeh Salami; Gholamhosein Aliahmmad

Zahedan Journal of Research in Medical Sciences

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Feasibility Study of Anti-Neurofilament Antibodies Detection by Indirect Quenching Fluoroimmunoassay

Author(s):

Alireza Nakhaee^1,*,

Manuchehr Messripour²,

Alireza Khosravi³,

Saeedeh Salami⁴,

Gholamhosein Aliahmmad⁵

1Zahedan University of Medical Sciences , Zahedan, Andorra

2Department of Biochemistry Isfahan Branch, Islamic Azad University Khorasgan, Isfahan, Iran

3ist , Zahedan University of Medical Sciences, Zahedan, Iran

4Department of Clinical Biochemistry, Research Center of Cellular and Molecular Biology, Zahedan University of Medical Sciences , Zahedan, Iran

5MSc of Sport and Rehabilitation Sciences, Faculty of Rehabilitation Sciences, Zahedan University of Medical Sciences, Zahedan, Iran

Zahedan Journal of Research in Medical Sciences:Vol. 16, issue 4; 11-44

Published online:Jan 07, 2012

Article type:Research Article

Received:Mar 16, 2015

Accepted:Oct 24, 2012

How to Cite:Alireza NakhaeeManuchehr MessripourAlireza KhosraviSaeedeh SalamiGholamhosein AliahmmadFeasibility Study of Anti-Neurofilament Antibodies Detection by Indirect Quenching Fluoroimmunoassay.Zahedan J Res Med Sci.16(4):11-44.

Abstract

Background: Neurofilaments (NFs) are the main constitutes of intermediate filaments in neurons. They are composed of three subunits with heavey, medium and low molecular weight. Anti-neurofilament antibodies exist in serum of patients with some neurodegenerative diseases.

Materials and Methods: A fluoroimmunoassay has been developed for determining of antibodies against neurofilaments, using an anti-fluorescein serum and fluorescein-labeled NFs. Antibodies raised against bovine spinal cord NFs in rabbit and the labeled NFs are incubated with anti-fluorescein serum at room temperature.

Results: At high levels, binding of anti-neurofilaments (anti-NFs) to labeled NFs prevented subsequent binding of the anti-fluorescein to fluorescein groups, resulting in little change in the signals of the label. Conversely, at low level of anti-NFs the free fraction of the labeled NFs is available to be bound by anti-fluorescein, which markedly reduced fluorescence intensity of label. Thus, the fluorescence intensity of assay mixture directly reflects the amount of anti-NFs antibodies in the serum.

Conclusion: It is concluded that the availability of fluorescein-labeled NFs and antibody directed against fluorescein group permit measurement of anti-NFs antibodies in serum of neurodegenerative patients.

Keywords

Neurofilament

Anti-Neurofilament

Fluoroimmunoassay

Neurodegenerative Disease

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