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

authors:

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

Zahedan University of Medical Sciences , Zahedan, Andorra
Department of Biochemistry Isfahan Branch, Islamic Azad University Khorasgan, Isfahan, Iran
ist , Zahedan University of Medical Sciences, Zahedan, Iran
Department of Clinical Biochemistry, Research Center of Cellular and Molecular Biology, Zahedan University of Medical Sciences , Zahedan, Iran
MSc 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: January 07, 2012
article type: Research Article
received: March 16, 2015
accepted: October 24, 2012

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. 

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.

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