Cloning and expression of an anti-nerve growth factor (NGF) antibody for studies using the neuroantibody approach.
1. The neuroantibody approach, based on the expression of selected monoclonal antibodies by cells of the nervous system, has recently been described (Cattaneo and Neuberger, 1987; Piccioli et al., 1991). In order to apply this experimental strategy to study the role of nerve growth factor (NGF) in the central nervous system (CNS), we exploited the monoclonal antibody (mAb), alpha D11, which neutralizes very efficiently the biological activity of NGF, both in vitro and in vivo (Cattaneo et al., 1988). 2. The alpha D11 antibody chains were cloned and expressed in COS cells as rat/human chimaeric proteins. The cloned antibody was shown to display all the properties of the parental alpha D11 antibody, including its ability to neutralize NGF biological activity. 3. This will allow us to engineer the expression of recombinant alpha D11 antibodies in the CNS, to study the role of NGF in the developing and adult nervous system. This approach can be extended to other neurotrophic factors for which neutralizing monoclonal antibodies are available.
Ruberti
F
F
CNR Institute of Neurobiology, Rome, Italy.
Bradbury
A
A
Cattaneo
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006825:Hybridomas / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000037:antagonists & inhibitors / Q000276:immunology* / Q000502:physiology*; D009424:Nervous System Physiological Phenomena; D016716:PC12 Cells; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000494:pharmacology; D015183:Restriction Mapping; D014162:Transfection
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D009414:Nerve Growth Factors; D012333:RNA, Messenger; D011994:Recombinant Proteins