Method for the manufacture of recombinant trypsin

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

Reexamination Certificate

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C435S006120, C435S069700, C435S069900

Reexamination Certificate

active

10181808

ABSTRACT:
The present invention relates to a method for the manufacture and purification of recombinant trypsinogen and trypsin inE. coliand yeast, using high yield expression vectors with and without secretion leader sequences. The invention further relates to an improved method and apparatus for carrying out protein refolding specifically useful for processing trypsinogen that has accumulated intracellularly in the form of inclusion bodies.

REFERENCES:
patent: 5521086 (1996-05-01), Scott et al.
patent: WO 97/00316 (1997-01-01), None
patent: WO 99/18219 (1999-04-01), None
Emi, M. et al, Cloning, characterization and nucleotide sequences of two cDNAs encoding human pancreatic trypsinogen. Gene 41: 305-310 (1986).
Bouffard, G.G. et al. A Collection of 1814 Human Chromosome 7-Specific STSs. Genome Research 7: 59-64 (1997).
Brodrick, J.W. et al. Human Cationic Trypsinogen: Purification, Characterization, and Characteristics of Autoactivation. The Journal of Biological Chemistry 253(8): 2732-2736 (1978).
Kay, J., et al., “The Autoactivation of Trypsinogen”, The Journal of Biological Chemistry, vol. 246, No. 21, 1971, pp. 6661-6665, XP-001030933.
Sahin-Toth, M., et al., “High-Affinity CA2+Binding Inhibits Autoactivation of Rat Trypsinogen”, Biochemical and Biophysical Research Communications, vol. 275, No. 2, Aug. 28, 2000, pp. 668-671, XP-002181528.

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