Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/130407
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Type: Journal article
Title: Development of mature BDNF-specific sandwich ELISA
Author: Lim, Y.
Zhong, J.H.
Zhou, X.F.
Citation: Journal of Neurochemistry, 2015; 134(1):75-85
Publisher: Wiley
Issue Date: 2015
ISSN: 0022-3042
1471-4159
Statement of
Responsibility: 
Yoon Lim, Jin-Hua Zhong, Xin-Fu Zhou
Abstract: Mature brain-derived neurotrophic factor (mBDNF) plays a vital role in the nervous system, whereas proBDNF elicits neurodegeneration and neuronal apoptosis. Although current enzyme-linked immunosorbent assay (ELISA) has been widely used to measure BDNF levels, it cannot differentiate mBDNF from proBDNF. As the function of proBDNF differs from mBDNF, it is necessary to establish an ELISA assay specific for the detection of mBDNF. Therefore, we aimed to establish a new mBDNF-specific sandwich ELISA. In this study, we have screened and found a combination of antibodies for a sandwich ELISA. A monoclonal antibody and sheep anti-BDNF were chosen as capture and detection antibody for sandwich ELISA respectively. The new ELISA showed no cross-reactivity to human recombinant NT-3, NT-4, nerve growth factor and negligible cross-reactivity (0.99-4.99%) for proBDNF compared to commercial ELISA kits (33.18-91.09%). The application of the new mBDNF ELISA was shown through the measurement of mBDNF levels in different brain regions of rats and in the brain of β-site amyloid precursor protein cleaving enzyme 1 (BACE1)(-/-) and WT mice and compared to western blot. Overall, this new ELISA will be useful for the measurement of mBDNF levels with high specificity. As the function of proBDNF differs from mBDNF (mature BDNF), it is necessary to establish an ELISA specific for the detection of mBDNF. Here, we present a novel sandwich ELISA which detects mBDNF with high specificity. This new ELISA will be useful for the measurement of mBDNF levels with high specificity in various human and animal tissues. proBDNF, precursor of BDNF; BDNF, brain-derived neurotrophic factor; NT-3, neurotrophin-3; NT-4, neurotrophin-4; NGF, nerve growth factor.
Keywords: ELISA and cross-reactivity; mature BDNF (mBDNF); proBDNF
Rights: © 2015 International Society for Neurochemistry.
RMID: 1000033755
DOI: 10.1111/jnc.13108
Grant ID: http://purl.org/au-research/grants/nhmrc/0595937
http://purl.org/au-research/grants/nhmrc/0480423
http://purl.org/au-research/grants/nhmrc/1017711
Appears in Collections:Biochemistry publications

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