Purification of a growth factor related to platelet-derived growth factor and a type beta transforming growth factor secreted by mouse neuroblastoma cells. A general strategy for the purification of basic polypeptide growth factors

A general strategy was developed for the purification of basic polypeptide growth factors. This method is a combination of gel filtration, weak-cation-exchange h.p.l.c. and reverse-phase h.p.l.c., separating the proteins according to size, charge and hydrophobicity respectively. All steps are carrie...

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Bibliographic Details
Published in:Biochemical journal 1989-01, Vol.257 (2), p.375-382
Main Authors: Van den Eijnden-Van Raaij, A J, Koornneef, I, Van Oostwaard, T M, Feyen, A, Kruijer, W, De Laat, S W, Van Zoelen, E J
Format: Article
Language:English
Subjects:
Animals
Blood Platelets - metabolism
Chromatography, Gel
Chromatography, High Pressure Liquid
Gene Expression Regulation
Growth Substances - isolation & purification
Growth Substances - metabolism
Mice
Neuroblastoma - analysis
Neuroblastoma - genetics
neuroblastoma cells
Peptides - metabolism
platelet-derived growth factor
Platelet-Derived Growth Factor - metabolism
platelets
transforming growth factor- beta
Transforming Growth Factors - metabolism
Tumor Cells, Cultured - metabolism
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Summary:A general strategy was developed for the purification of basic polypeptide growth factors. This method is a combination of gel filtration, weak-cation-exchange h.p.l.c. and reverse-phase h.p.l.c., separating the proteins according to size, charge and hydrophobicity respectively. All steps are carried out at low pH with exclusively volatile acidic buffer solutions. The sterile conditions and easy removal of salt by freeze-drying facilitate the detection of the growth factors by biological assays. By using this method, homogeneous preparations of two basic growth factors were purified in high yield from mouse-neuroblastoma-Neuro-2A-cell-conditioned medium. It is shown that these purified factors are biochemically and immunologically related to platelet-derived growth factor and type beta transforming growth factor from human platelets.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj2570375