Development of a Novel Highly Sensitive mBDNF ELISA
PRESENTER:
Dr. Naomi Tsurutani | Sales Manager at FUJIFILM Wako Chemicals U.S.A. Corporation
PRESENTER'S BIO:
Naomi Tsurutani received her Ph.D. in Virology/Immunology from Tokyo Medical and Dental University in Tokyo, Japan. Before joining FUJIFILM Wako, she worked as an immunology researcher at Emory University, UMass Medical School, and UConn Health.
At FUJIFILM Wako, she helps researchers better perform and communicate their research using FUJIFILM Wako products.
ABSTRACT:
BDNF, a member of the neurotrophin family of growth factors, is involved in neurogenesis and synaptogenesis and plays critical roles in numerous signaling pathways associated with nervous system disorders ranging from depression, autism, and schizophrenia in addition to heart disease, diabetes, gout, periodontal disease, stress, and cognitive effects associated with exercise. BDNF is synthesized as the precursor proBDNF, which undergoes intra or extracellular cleavage to produce mature BDNF (mBDNF). Being able to distinguish between the two forms is imperative, as both proBDNF and mBDNF are found in the brain and the periphery and exert opposite effects by binding to the p75 neurotrophin receptor and tyrosine kinase receptor B receptors, respectively. However, most commercially available BDNF ELISA kits fail to specifically detect mBDNF in mouse serum and plasma, compromising studies targeting these proteins. To address this problem, we used an N-terminal end-specific anti-mBDNF antibody to develop a highly sensitive ELISA. The new ELISA showed very low reactivity (1.30%) to proBDNF and other NGF family proteins (human NGF, NT-3 and NT-4).
Moreover, by using biotin-labeled, streptavidin-conjugated horseradish peroxidase and a chemiluminescent substrate, we achieved a high sensitivity of 0.116 pg/ml. The ELISA was tested using brain lysates from BDNF KO mice, resulting in deficient levels of mBDNF detection compared to wild-type mice lysates [≈500 pg/mL]. This indicates that our new ELISA was superior in sensitivity and specificity to other commercially available conventional methods. The new ELISA also showed good recovery rates (84.8-110%) from mouse, rat, and human plasma and serum samples in dilution linearity and spike-recovery tests. Furthermore, mBDNF could be detected in the serum of wild-type [6.96 - 9.01 pg/mL] and autism model mice [12.4-18.1 pg/mL], and human saliva samples [0.296 - 4.09 pg/mL], the latter being a result that other conventional methods have not yet achieved. In conclusion, our novel ELISA is expected to be a valuable tool in BDNF basic research and mental diseases research.
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