Bifunctional fusion between nerve growth factor and a transferrin receptor antibody.
The cDNAs encoding the variable regions of the heavy and light chains of a murine antibody specific for the human TfR were cloned and a human chimera (gamma1, kappa) was produced. A gene fusion was created by joining the 3' end of the coding region of the human nerve growth factor (NGF) precursor to the 5' end of the heavy chain variable region of the chimeric antibody. When expressed with the unmodified light chain in mammalian cells, the protein fusion is properly processed, assembled, and secreted. Subsequent purification and characterization established the uncompromised bifunctional activities of the protein, relative to the unmodified components, as demonstrated by its ability to both bind to the human TfR and induce neurite outgrowth in primary sympathetic or spinal ganglia and in trkA-transfected pheochromocytoma cells. The ability to generate biologically active NGF fused to a TfR targeting antibody, which was previously shown to cross the blood-brain barrier, may offer a novel way to deliver NGF and other neurotrophic factors to the central nervous system.
McGrath
J P
JP
Alkermes, Inc., Cambridge, Massachusetts 02139, USA.
Cao
X
X
Schutz
A
A
Lynch
P
P
Ebendal
T
T
Coloma
M J
MJ
Morrison
S L
SL
Putney
S D
SD
D000818:Animals; D000918:Antibody Specificity; D002678:Chimera / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification*; D005815:Genetic Code; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis*; D007135:Immunoglobulin Variable Region; D051379:Mice; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000096:biosynthesis / Q000235:genetics*; D011990:Receptors, Transferrin / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D018076:DNA, Complementary; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D009414:Nerve Growth Factors; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins