Large-Scale Purification of Recombinant Human Angiostatin

A process for the purification of recombinant human angiostatin (rhAngiostatin), produced by Pichia pastoris fermentation operated at the 2000-L scale, is reported. rhAngiostatin was recovered and purified directly from crude fermentation broth by cation exchange expanded bed adsorption chromatograp...

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Bibliographic Details
Published in:Protein expression and purification 2000-11, Vol.20 (2), p.216-227
Main Authors: Shepard, Scot R., Boucher, Robert, Johnston, Jeremy, Boerner, Renee, Koch, George, Madsen, John W., Grella, Davida, Sim, B.Kim Lee, Schrimsher, Jeffrey L.
Format: Article
Language:English
Subjects:
Ammonium Sulfate - metabolism
Angiostatins
Antineoplastic Agents - isolation & purification
Chromatography - methods
Chromatography, High Pressure Liquid
Durapatite - metabolism
Electrophoresis, Polyacrylamide Gel
Humans
Peptide Fragments - biosynthesis
Peptide Fragments - genetics
Peptide Fragments - isolation & purification
Phosphates
Pichia - genetics
Plasminogen - biosynthesis
Plasminogen - genetics
Plasminogen - isolation & purification
Recombinant Proteins - biosynthesis
Recombinant Proteins - isolation & purification
Reproducibility of Results
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Summary:A process for the purification of recombinant human angiostatin (rhAngiostatin), produced by Pichia pastoris fermentation operated at the 2000-L scale, is reported. rhAngiostatin was recovered and purified directly from crude fermentation broth by cation exchange expanded bed adsorption chromatography. Anion exchange chromatography, hydroxyapatite chromatography, and hydrophobic interaction chromatography were used for further purification. Full-length rhAngiostatin was separated from rhAngiostatin molecules fragmented by endoproteolysis. On average, 140 g of rhAngiostatin was produced per batch, with an overall yield of 59% (n = 9). The purification process was completed in approximately 48 h and used only inexpensive and nontoxic raw materials. Methods development, process synthesis, and process scale-up data are presented and discussed.
ISSN:1046-5928
1096-0279
DOI:10.1006/prep.2000.1276