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

AUTHORS

Alireza Nakhaee 1 , * , Manuchehr Messripour 2 , Alireza Khosravi 3 , Saeedeh Salami 4 , Gholamhosein Aliahmmad 5

1 Zahedan University of Medical Sciences , Zahedan, Andorra

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

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

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

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

How to Cite: Nakhaee A , Messripour M, Khosravi A, Salami S, Aliahmmad G. Feasibility Study of Anti-Neurofilament Antibodies Detection by Indirect Quenching Fluoroimmunoassay, Zahedan J Res Med Sci. 2014 ; 16(4):11-44.

ARTICLE INFORMATION

Zahedan Journal of Research in Medical Sciences: 16 (4); 11-44
Published Online: January 07, 2012
Article Type: Research Article
Received: March 16, 2015
Accepted: October 24, 2012

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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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