Purification and identification of brain-derived neurotrophic factor from human serum.
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.
Amgen Center, Thousand Oaks, California 91320, USA.
R D
RD
Rosenfeld
L
L
Zeni
M
M
Haniu
J
J
Talvenheimo
S F
SF
Radka
L
L
Bennett
J A
JA
Miller
A A
AA
Welcher
D000595:Amino Acid Sequence; D001923:Brain Chemistry; D019208:Brain-Derived Neurotrophic Factor; D002498:Centrifugation; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D009419:Nerve Tissue Proteins / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D011145:Polyvinyls; D005465:Fluorocarbon Polymers
D019208:Brain-Derived Neurotrophic Factor; D008567:Membranes, Artificial; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers