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Purification and characterization of fab fragments from anti‐mouse NGF polyclonal antibodies
Author(s) -
Callegaro L.,
Skaper S. D.,
Vantini G.,
Benvegnù D.,
Di Martino A.,
Schiavo N.,
Triban C.,
Minozzi C.,
Leon A.
Publication year - 1990
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.300030503
Subject(s) - polyclonal antibodies , nerve growth factor , antibody , dorsal root ganglion , in vitro , affinity chromatography , central nervous system , biology , in vivo , nervous system , sepharose , microbiology and biotechnology , chemistry , biochemistry , immunology , neuroscience , enzyme , spinal cord , receptor
A functional role for Nerve Growth Factor (NGF) in the peripheral nervous system is well‐documented, but a similar case for NGF in the central nervous system remains to be established. One approach to answering this question would be the availability of high‐affinity monospecific Fab fragments obtained against NGF. In the present studies we describe the preparation and characterization of such Fab fragments from anti‐mouse NGF polyclonal antibodies. Following their purification by the use of a NGF Sepharose‐coupled affinity column, the Fab fragments were examined for biological competence in several ways. In vitro , the anti‐Fab fragments blcoked the neuronotrophic activity of NGF, as measured by the survival of chicken embryonic day 8 dorsal root ganglion neurons. In vivo , these Fab fragments, when administered systemically to neonatal rats, produced a decrease of noradrenaline levels in two sympathetically innervated organs, the heart and the spleen. These findings suggest that affinity purified Fab fragments of anti‐NGF antibodies can be a useful tool for studying the physiological function of NGF in the nervous system.