Recombinant human nerve growth factor for clinical trials: protein expression, purification, stability and characterisation of binding to infusion pumps.
Nerve growth factor (NGF) has been suggested to be of therapeutic benefit to patients with Alzheimer's disease. One of the early changes in this disease is a loss of cholinergic function within the brain, and NGF is able to rescue cholinergic neurons both in vitro and in vivo. We describe the production of recombinant human beta-NGF (rhNGF), using baculovirus infection of insect cells; its purification, formulation and subsequent stability for use in clinical trials. Tests were also carried out to monitor release of protein from infusion pumps and catheters for intracerebroventricular administration (icv). Initial problems with non-specific binding were overcome using a blocking formula.
Molecular Neurobiology Unit, University Research Centre for Neuroendocrinology (Care of the Elderly), University of Bristol, BS2 8HW, Bristol, UK. shelley.allen@bristol.ac.uk
S J
SJ
Allen
A G
AG
Robertson
S J
SJ
Tyler
G K
GK
Wilcock
D
D
Dawbarn
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001681:Biological Assay; D002460:Cell Line; D002986:Clinical Trials as Topic; D006168:Guinea Pigs; D006801:Humans; D007260:Infusion Pumps; D051379:Mice; D020932:Nerve Growth Factor / Q000302:isolation & purification* / Q000627:therapeutic use / Q000633:toxicity; D016716:PC12 Cells; D011485:Protein Binding; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000627:therapeutic use / Q000633:toxicity; D018411:Spodoptera
D011994:Recombinant Proteins; D020932:Nerve Growth Factor