PURIFICATION AND IDENTIFICATION OF BRAIN-DERIVED NEUROTROPHIC FACTOR FROM HUMAN SERUM

被引:147
作者
ROSENFELD, RD [1 ]
ZENI, L [1 ]
HANIU, N [1 ]
TALVENHEIMO, J [1 ]
RADKA, SF [1 ]
BENNETT, L [1 ]
MILLER, JA [1 ]
WELCHER, AA [1 ]
机构
[1] SUPRAGEN INC,LAKEWOOD,CO 80214
关键词
D O I
10.1006/prep.1995.1062
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Brain-derived neurotrophic factor (BDNF), a 27-kDa noncovalently linked homodimer with subunits of approximate to 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. (C) 1995 Academic Press, Inc.
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页码:465 / 471
页数:7
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