Process for the preparation of a new-type active substance selec

Details Process for the preparation of a new-type active substance selec Process for the preparation of a new-type active substance selec

1984-07-19

1986-05-13

Rosen, Sam

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435 68, 424101, 514 2, 514 8, 530380, 530414, 530830, C12P 2106

Patent

active

045886857

ABSTRACT:
The invention involves isolating a food intake suppressant by subjecting blood serum to ultrafiltration transmitting up to a molecular weight of 30,000 daltons, partially evaporating the filtrate, removing the insoluble part from the concentrate obtained, adding trichloroacetic acid up to a concentration of 5 to 25 weight/volume percent to the liquid phase at a temperature of 0.degree. to 10.degree. C., removing the proteins precipitated, subjecting the obtained solution to chromatography on a gel with a void volume below a molecular weight of 4000 daltons, eluting with a solution of pH 6.0 to 7.0, concentrating the biologically active fractions, chromatographing again on a gel with a void volume below a molecular weight of 4000 daltons, fractionating by elution with water, lyophilizing the active fractions, dissolving the lyophilized fractions in a buffer of pH 8.1 to 8.2, adding trypsin and chymotrypsin to the solution in a substantially identical amount of 0.01 to 0.2 by weight as calculated for the total weight of the lyophilized product, subjecting the mixture to digestion at a temperature of 36.degree. C. to 39.degree. C., preferably with occasionally shaking from 20 to 30 hours, adding after the first 5 hours trypsin and chymotrypsin in a substantially half amount as calculated for the original enzyme quantity and continuing the digestion, then adding trichloroacetic acid to the reaction mixture up to a concentration of 4 to 6 weight/volume percent at a temperature between 0.degree. and 10.degree. C., keeping the mixture at a temperature between -15.degree. C. and 5.degree. C. for 1 to 24 hours, removing the insoluble part, subjecting the reaction mixture to chromatography on a gel with a void volume below a molecular weight of 4000 daltons, fractionating by elution with water and lyophilizing the biologically active fractions.

REFERENCES:
patent: 4294825 (1981-10-01), Knoll et al.
patent: 4430264 (1984-02-01), Knoll et al.

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