Purification and characterization of Fab fragments from anti-mouse NGF polyclonal antibodies.
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 blocked 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.
Callegaro
L
L
Fidia Research Laboratory, Abano Terme, Italy.
Skaper
S D
SD
Vantini
G
G
Benvegnù
D
D
Di Martino
A
A
Schiavo
N
N
Triban
C
C
Minozzi
C
C
Leon
A
A
D000818:Animals; D002642:Chick Embryo; D005260:Female; D015151:Immunoblotting; D007140:Immunoglobulin Fab Fragments / Q000008:administration & dosage / Q000276:immunology / Q000302:isolation & purification*; D008297:Male; D051379:Mice; D009414:Nerve Growth Factors / Q000276:immunology* / Q000302:isolation & purification; D009638:Norepinephrine / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D007140:Immunoglobulin Fab Fragments; D009414:Nerve Growth Factors; D009638:Norepinephrine