Purification and refolding of recombinant proteins

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Lymphokines – e.g. – interferons – interlukins – etc.

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530350, 530399, 530303, 530418, 530420, 530427, 435 6952, C07K 324, C07K 1506, C07K 740, C07K 1300

Patent

active

049239678

ABSTRACT:
The present invention discloses a new method for solubilizing and refolding recombinant proteins expressed as granules. The method involves sulfitolysis and the formation of a precipitate of protein-S-sulfonate by warming. The precipitate has been found to contain protein in high purity. In addition, proper folding takes place if the desired protein is fully reduced and passed through an intermediate concentration of denaturant which allows for a transition between its folded and unfolded states.

REFERENCES:
patent: 4569790 (1986-02-01), Koths et al.
patent: 4656249 (1987-04-01), Tregear et al.
Tsugawa, R., Production of Recombinant Interleukin-2, Bio-Fair Tokyo '86.
Kato, K. et al., Purification and Characterization of Human Interleukin-2 Produced in Escherichia coli, Biochem. Biophys. Res. Comm. 130:692 (1985).
Weir, M. P., Purification and Renaturation of Recombinant Human Interleukin-2, Biochem. J. 245:85 (1987).
Furman, T. C., et al., Recombinant Human Insulin-Like Growth Factor II Expressed in Escherichia coli, Bio/Technology, 5:1047 (1987).
Weir, M. P., et al., Micropreparative Purification of Recombinant Human Interleukin-2, Journal of Chromatography, 396:209 (1987).
Tsuji, et al., Characterization of Disulfide Bonds in Inclusion Bodies and Its Oxidative Refolding, Biochemistry, 26:3129 (1987).

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