Recombinant human nerve growth factor with a marked activity in vitro and in vivo.
Recombinant human nerve growth factor (rhNGF) is regarded as the most promising therapy for neurodegeneration of the central and peripheral nervous systems as well as for several other pathological conditions involving the immune system. However, rhNGF is not commercially available as a drug. In this work, we provide data about the production on a laboratory scale of large amounts of a rhNGF that was shown to possess in vivo biochemical, morphological, and pharmacological effects that are comparable with the murine NGF (mNGF), with no apparent side effects, such as allodynia. Our rhNGF was produced by using conventional recombinant DNA technologies combined with a biotechnological approach for high-density culture of mammalian cells, which yielded a production of approximately 21.5 +/- 2.9 mg/liter recombinant protein. The rhNGF-producing cells were thoroughly characterized, and the purified rhNGF was shown to possess a specific activity comparable with that of the 2.5S mNGF by means of biochemical, immunological, and morphological in vitro studies. This work describes the production on a laboratory scale of high levels of a rhNGF with in vitro and, more important, in vivo biological activity equivalent to the native murine protein.
Colangelo
Anna M
AM
Laboratory of Neuroscience R. Levi-Montalcini, Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy.
Finotti
Nicoletta
N
Ceriani
Michela
M
Alberghina
Lilia
L
Martegani
Enzo
E
Aloe
Luigi
L
Lenzi
Laura
L
Levi-Montalcini
Rita
R
D000818:Animals; D000831:Animals, Newborn; D019149:Bioreactors; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002645:Chickens; D004789:Enzyme Activation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D009414:Nerve Growth Factors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D010766:Phosphorylation; D020917:Receptor, trkA / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
D009414:Nerve Growth Factors; D011994:Recombinant Proteins; D020917:Receptor, trkA