Recombinant human BDNF protein

Summary

• High purity human BDNF protein (Uniprot: P23560)
• 14 kDa
• >98%, by SDS-PAGE quantitative densitometry
• Expressed in E. coli
• Animal origin-free (AOF) and carrier protein-free
• Manufactured in Qkine's Cambridge, UK laboratories
• Lyophilized from acetonitrile, TFA
• Resuspend in sterile-filtered water at >50 µg/ml, add carrier protein if desired, prepare single use aliquots and store frozen at -20 °C (short-term) or -80 °C (long-term).

Featured applications

• Differentiation of midbrain dopaminergic neurons
• Differentiation of iPSC derived neural progenitors to neurons

Qkine BDNF was more bioactive than BDNF from an alternative supplier. Recombinant BDNF activity was determined using a NFAT-luciferase reporter assay in transfected HEK293T cells also expressing TrkB. Cells are treated in triplicate with a serial dilution of Qkine BDNF (Qk050, green) or alternative supplier BDNF (Supplier B, black) for 6 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. Data from Qk050 lot #104465.

Protein background

Brain-derived neurotrophic factor (BDNF) is a member of neurotrophin family which includes nerve growth factor (NGF) and neurotrophin–3/4/5 (NT–3/4/5) [1,2]. BDNF plays a crucial role during embryonic development and the maintenance of the nervous system during adulthood. It mediates many aspects, including the survival, growth, maintenance, and differentiation of neural stem cells and mature neurons [3,4]. It stimulates neurogenesis and promotes the growth of dendrites and axons [5,6]. It is also involved in synaptic plasticity, forming and strengthening synapses in response to experience and learning [7]. Finally, it regulates the neuroprotection as it is involved in neural repair and recovery [8].

In cell culture, recombinant human BDNF supports the survival and growth of neurons and establishes and strengthens synapses. It is used in combination with other growth factors to maintain neurons and to differentiate human-induced pluripotent stem cell-derived neural progenitors into neurons [5]. It is used for the differentiation and maturation of dopaminergic neurons with glial-derived neurotrophic factor (GDNF), FGF-8a/FGF-8b, and Shh [9–11]. Glutamatergic and GABAergic neurons are maintained with combinations of BDNF, GDNF, NT-3, or insulin-like growth factor 1 (IGF-1) [12–14]. Cholinergic neurons are obtained using BDNF, IGF-1, and NGF [15,16]. Also, the culture of cortical organoids require BDNF and GDNF for maturation [17].

BDNF is released by various cells including neurons, glial cells (astrocytes and microglia), immune cells (T cells and macrophages), as well as skeletal muscles. It has several precursor isoforms and its mature active form is a dimer stabilised by a cysteine knot composed of 119 amino acids and a molecular weight of around 13 kDa [18]. The mature isoform binds to the receptor tropomyosin receptor kinase B (TrkB) with high affinity to activate MAPK, PI3K and PLC-γ signalling cascades [19,20]. These pathways are related to the survival of neurons, growth of dendrites and axons, development of synapses, and learning- and memory-dependent synaptic plasticity19. In addition, this protein binds to p75 neurotrophin receptor (p75NTR) with lower affinity to activate the NF-kB activation [21].

Impaired BDNF signaling has been linked to several diseases such as neurodegenerative diseases such as Alzheimer’s and Parkinson’s diseases, neurological disorders such as depression and anxiety disorders, as well as spinal cord injury [22,23]. Recombinant BDNF and other neurotrophins such as NGF and NT-3 are a growing area of research as potential targets [22,24]. For example, pharmacological agents or gene therapy could increase BDNF production and release which may have therapeutic potential for conditions like amyotrophic lateral sclerosis and depression. Also, targeted delivery of this protein could be beneficial to preserve nerve function, promote healthy brain ageing, and regenerate the loss of neurons.

Background references

1. Maisonpierre, P. C. et al. NT-3, BDNF, and NGF in the developing rat nervous system: Parallel as well as reciprocal patterns of expression. Neuron 5, 501–509 (1990). doi: 10.1016/0896-6273(90)90089-x
2. Skaper, S. D. The Neurotrophin Family of Neurotrophic Factors: An Overview. in Neurotrophic Factors: Methods and Protocols (ed. Skaper, S. D.) 1–12 (Humana Press, 2012). doi:10.1007/978-1-61779-536-7_1.
3. Ghosh, A., Carnahan, J. & Greenberg, M. E. Requirement for BDNF in Activity-Dependent Survival of Cortical Neurons. Science 263, 1618–1623 (1994). doi: 10.1126/science.7907431
4. Hyman, C. et al. BDNF is a neurotrophic factor for dopaminergic neurons of the substantia nigra. Nature 350, 230–232 (1991). doi: 10.1038/350230a0
5. Brafman, D. A. Generation, Expansion, and Differentiation of Human Pluripotent Stem Cell (hPSC) Derived Neural Progenitor Cells (NPCs). in Stem Cell Renewal and Cell-Cell Communication: Methods and Protocols (ed. Turksen, K.) 87–102 (Springer, 2015). doi:10.1007/7651_2014_90
6. Chen, B.-Y. et al. Brain-derived neurotrophic factor stimulates proliferation and differentiation of neural stem cells, possibly by triggering the Wnt/β-catenin signaling pathway. J. Neurosci. Res. 91, 30–41 (2013). doi: 10.1002/jnr.23138
7. Bramham, C. R. & Messaoudi, E. BDNF function in adult synaptic plasticity: The synaptic consolidation hypothesis. Prog. Neurobiol. 76, 99–125 (2005). doi: 10.1016/j.pneurobio.2005.06.003
8. Park, H. & Poo, M. Neurotrophin regulation of neural circuit development and function. Nat. Rev. Neurosci. 14, 7–23 (2013). doi: 10.1038/nrn3379
9. Perrier, A. L. et al. Derivation of midbrain dopamine neurons from human embryonic stem cells. Proc. Natl. Acad. Sci. U. S. A. 101, 12543–12548 (2004). doi: 10.1073/pnas.0404700101
10. Brennand, K. et al. Modeling schizophrenia using hiPSC neurons. Nature 473, 221–225 (2011). doi: 10.1038/nature09915
11. Kriks, S. et al. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease. Nature 480, 547–551 (2011). doi: 10.1038/nature10648
12. Fernandopulle, M. S. et al. Transcription-factor mediated differentiation of human iPSCs into neurons. Curr. Protoc. Cell Biol. 79, e51 (2018). doi: 10.1002/cpcb.51
13. Li, X.-J. et al. Specification of motoneurons from human embryonic stem cells. Nat. Biotechnol. 23, 215–221 (2005). doi: 10.1038/nbt1063
14. Lin, L., Yuan, J., Sander, B. & Golas, M. M. In Vitro Differentiation of Human Neural Progenitor Cells Into Striatal GABAergic Neurons. Stem Cells Transl. Med. 4, 775–788 (2015). doi: 10.5966/sctm.2014-0083
15. Liu, Y. et al. Medial ganglionic eminence–like cells derived from human embryonic stem cells correct learning and memory deficits. Nat. Biotechnol. 31, 440–447 (2013). doi: 10.1038/nbt.2565
16. Nilbratt, M., Porras, O., Marutle, A., Hovatta, O. & Nordberg, A. Neurotrophic factors promote cholinergic differentiation in human embryonic stem cell-derived neurons. J. Cell. Mol. Med. 14, 1476–1484 (2010). doi: 10.1111/j.1582-4934.2009.00916.x
17. Jacob, F. et al. Human Pluripotent Stem Cell-Derived Neural Cells and Brain Organoids Reveal SARS-CoV-2 Neurotropism Predominates in Choroid Plexus Epithelium. Cell Stem Cell 27, 937-950.e9 (2020). doi: 10.1016/j.stem.2020.09.016
18. Barde, Y. a., Edgar, D. & Thoenen, H. Purification of a new neurotrophic factor from mammalian brain. EMBO J. 1, 549–553 (1982). doi: 10.1002/j.1460-2075.1982.tb01207.x
19. Colucci-D’Amato, L., Speranza, L. & Volpicelli, F. Neurotrophic Factor BDNF, Physiological Functions and Therapeutic Potential in Depression, Neurodegeneration and Brain Cancer. Int. J. Mol. Sci. 21, 7777 (2020). doi: 10.3390/ijms21207777
20. Foltran, R. B. & Diaz, S. L. BDNF isoforms: a round trip ticket between neurogenesis and serotonin? J. Neurochem. 138, 204–221 (2016). doi: 10.1111/jnc.13658
21. Reichardt, L. F. Neurotrophin-regulated signalling pathways. Philos. Trans. R. Soc. B Biol. Sci. 361, 1545–1564 (2006). doi: 10.1098/rstb.2006.1894
22. Mattson, M. P., Maudsley, S. & Martin, B. BDNF and 5-HT: a dynamic duo in age-related neuronal plasticity and neurodegenerative disorders. Trends Neurosci. 27, 589–594 (2004). doi: 10.1016/j.tins.2004.08.001
23. Mattson, M. P. & Scheff, S. W. Endogenous Neuroprotection Factors and Traumatic Brain Injury: Mechanisms of Action and Implications for Therapy. J. Neurotrauma 11, 3–33 (1994). doi: 10.1089/neu.1994.11.3
24. Keefe, K. M., Sheikh, I. S. & Smith, G. M. Targeting Neurotrophins to Specific Populations of Neurons: NGF, BDNF, and NT-3 and Their Relevance for Treatment of Spinal Cord Injury. Int. J. Mol. Sci. 18, 548 (2017). doi: 10.3390/ijms18030548

Publications using Recombinant human BDNF protein (Qk050)

• Human retinal ganglion cell neurons generated by synchronous BMP inhibition and transcription factor mediated reprogramming. Agarwal D et al.
  DOI: doi: 10.1038/s41536-023-00327-x
• Differentiation of RGC Induced Neurons (RGC-iNs).
  Agarwal D & Wahlin K
  DOI: dx.doi.org/10.17504/protocols.io.14egn2pqzg5d/v2
• A short sequence targets transmembrane proteins to primary cilia
  Macarelli V, Harding EC, Gershlick DC, Merkle FT.
  DOI: 10.3390/cells13131156
• Heterogeneity in oligodendrocyte precursor cell proliferation is dynamic and driven by passive bioelectrical properties
  Pivoňková H, Sitnikov S, Kamen Y et al.
  DOI: 10.1016/j.celrep.2024.114873
• Parkinson’s associated protein DJ-1 regulates intercellular communication via extracellular vesicles in oxidative stress
  Page T, Musi CA, Bakker SE et al.
  DOI: https://doi.org/10.21203/rs.3.rs-5669239/v1
• Profiling human hypothalamic neurons reveals a candidate combination drug therapy for weight loss
  Chen HJC, Yang A, Mazzaferro S et al.
  DOI: https://doi.org/10.1101/2023.07.18.549357
• Synchronous 3D patterning of diverse CNS progenitors generates motor neurons of broad axial identity
  Buchner F, Dokuzluoglu Z, Thomas J et al.
  DOI: https://doi.org/10.1101/2024.12.13.628384
• Neuronal differentiation enhances a cytoplasmic pool of tousled-like kinase 2 (TLK2)
  Nuhu-Soso L, Denton H, Goffin DL, Hahn I and Evans GJO.
  DOI: 10.1101/2025.02.20.639293
• Familial ALS/FTD-associated RNA-Binding deficient TDP-43 mutants cause neuronal and synaptic dysregulation in vitro
  Magarotto M, Gawne RT, Vilkaite G, Mason AS, Chen H-J
  DOI: doi.org/10.1101/2025.03.26.645507

FAQ

What is a recombinant brain-derived neurotrophic factor?
Recombinant Brain-Derived Neurotrophic Factor is a synthetic form of BDNF. Qkine recombinant human BDNF is expressed in E. coli.

What does BDNF do?
BDNF plays a crucial role during embryonic development and the maintenance of the nervous system during adulthood including the survival, growth, maintenance, and differentiation of neurons, growth of dendrites and axons, and synaptic plasticity.

What is the difference between NGF, GDNF and BDNF?
NGF, GDNF, and BDNF are all part of the neurotrophins or neurotrophic factors family. They all regulate the development, growth, and survival of neurons and have more specific roles. NGF is associated with the growth and maintenance of sensory and sympathetic neurons. BDNF and GDNF are both associated with dopaminergic, glutamatergic, and GABAergic neurons. In addition, BDNF plays a crucial role in synapses formation whereas GDNF is involved in the growth and maintenance of other glial cells such as astrocytes.

What is the BDNF gene?
The BDNF protein is encoded by the BDNF gene on chromosome 11.

What are examples of neurotrophic factors?
Neurotrophic factors (also known as neurotrophins) include Nerve Growth Factor (NGF), Brain-Derived Neurotrophic Factor (BDNF), Glial-Derived Neurotrophic Factor (GDNF), Neurotrophin-3 (NT-3), and Neurotrophin-4/5 (NT-4/5).

What do neurotrophin proteins do?
Neurotrophin proteins regulate neuronal growth and survival, neuroprotection, neurotransmitter release, synaptic function, and plasticity within the adult central and peripheral nervous system.

What is the role of neurotrophins in programmed cell death?
Neurotrophins promote the survival and growth of neurons by inhibiting apoptosis (programmed cell death) in a process called neuroprotection.