Purification and Identification of Brain-Derived Neurotrophic Factor from Human Serum

Amgen Center, Thousand Oaks, California 91320, USA.
Protein Expression and Purification (Impact Factor: 1.7). 09/1995; 6(4):465-71. DOI: 10.1006/prep.1995.1062
Source: PubMed


Brain-derived neurotrophic factor (BDNF), a 27-kDa noncovalently linked homodimer with subunits of approximately 13.5 kDa as viewed by SDS-PAGE, is thought to be primarily produced in the central nervous system. We report here the isolation of BDNF from pooled normal human sera, using a two-step purification process followed by SDS-PAGE, transfer to a polyvinylidene difluoride membrane, and subsequent identification of the protein by sequence analysis of the appropriate band(s) from the membrane. The level of BDNF in pooled human sera was estimated to be approximately 15 ng/ml as determined by an enzyme-linked immunosorbant assay. The average for six individuals was 18.9 +/- 5.7 ng/ml. There is an approximately 200-fold increase in the levels of BDNF in serum relative to plasma. Results from experiments using differential centrifugation suggest that the source of this increase is due to release from platelets. The presence of high levels of BDNF in serum suggests a role for this neurotrophin either in nerve repair at sites of injured tissue or in nonneuronal functions.

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    • "BDNF was first identified in 1982 by Yves Barde and Hans Thoenen [1], and its importance as a therapeutic agent in neuropsychiatric disorders [2] and in improving plasticity and cognition in healthy conditions has been well documented [3]. Rosenfeld et al. originally showed that BDNF could be detected in human serum or plasma [4]. Later, a positive correlation between brain and plasma levels of BDNF was found [5]. "
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    ABSTRACT: OBJECTIVES: Physical exercise up-regulates brain-derived neurotrophic factor (BDNF) in brain and blood. However, there is yet no consensus about the adequate blood processing conditions to standardize its assessment. We aimed to find a reliable blood sample processing method to determine changes in BDNF due to exercise. DESIGN AND METHODS: Twelve healthy university students performed an incremental cycling test to exhaustion. At baseline, immediately after exercise, and 30 and 60minutes of recovery, venous blood was drawn and processed under different conditions, i.e. whole blood, serum coagulated for 10minutes and 24hours, total plasma, and platelet-free plasma. BDNF concentration was measured by ELISA. RESULTS: Exercise increased BDNF in whole blood and in serum coagulated for 24hours when corrected by hemoconcentration. We did not find effects of exercise on BDNF in serum coagulated for 10minutes or in plasma samples. Plasma shows heterogeneous BDNF values in response to exercise that are not prevented when platelets are eliminated whilst homogeneous BDNF levels were found in whole blood or serum coagulated for 24hours samples. CONCLUSIONS: In exercise studies, BDNF levels should be adjusted by hemoconcentration. Our data highlight the importance of blood sample selection since the differences between each one affect significantly the BDNF factor changes due to exercise.
    Clinical Biochemistry 11/2014; 1(3). DOI:10.1016/j.clinbiochem.2014.11.013 · 2.28 Impact Factor
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    • "This was performed at Biochemistry Department laboratory, Cairo University. Our reference range was 18.9 ± 5.7 ng/ml (Rosenfeld et al., 1995). "
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    ABSTRACT: Stroke is a leading cause of functional impairments. High percentage of these patients will experience some degree of cognitive affection, ranging from mild cognitive impairment to dementia. Demonstrate the role of aerobic exercises enhancing cognitive functions and its effect on Brain Derived Neurotrophic factor (BDNF) in post-ischemic stroke patients in the territory of anterior circulation. We included thirty Egyptian ischemic stroke patients in the territory of anterior circulation. They were divided into 2 groups; group 1 (G1) were subjected to physiotherapy program without aerobic exercises and group 2 (G2) were subjected to the same previous program followed by aerobic exercises. Both groups were subjected to pre- and post-treatment Addenbrookes's Cognitive Examination- Revised (ACER) and serum level of BDNF. Our results showed a significant improvement in ACER score in G2 compared to G1 post-treatment (p = 0.017). BDNF serum level significantly increased in G2 post-treatment compared to pre-treatment (p = 0.001) and compared to G1 group (p = 0.0458). ACER improvement was positively correlated to increase in serum level of BDNF (r = 0.53, p = 0.044). Aerobic exercises improve cognitive functions of ischemic stroke patients. This improvement is related to the increase in serum level of BDNF.
    Neurorehabilitation 11/2013; 34(1). DOI:10.3233/NRE-131020 · 1.12 Impact Factor
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    • "Language was limited to English. The search covered the period from January 1995 to July 2012 since the first evidence for the presence of BDNF in human blood emerged in 1995 (Rosenfeld et al., 1995). Additionally, we searched the references of published studies manually. "
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    ABSTRACT: The purpose of this study was to summarize the effects of physical activity and exercise on peripheral brain-derived neurotrophic factor (BDNF) in healthy humans. Experimental and observational studies were identified from PubMed, Web of Knowledge, Scopus, and SPORT Discus. A total of 32 articles met the inclusion criteria. Evidence from experimental studies suggested that peripheral BDNF concentrations were elevated by acute and chronic aerobic exercise. The majority of the studies suggested that strength training had no influence on peripheral BDNF. The results from most observational studies suggested an inverse relationship between the peripheral BDNF level and habitual physical activity or cardiorespiratory fitness. More research is needed to confirm the findings from the observational studies.
    Scandinavian Journal of Medicine and Science in Sports 04/2013; 24(1). DOI:10.1111/sms.12069 · 2.90 Impact Factor
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