Polypeptides from urogastrone

Claim 1

A PROCESS FOR THE FRACTIONATION OF CRUDE UROGASTRONE INTO THREE COMPONENTS WHICH COMPRISES THE STEPS OF: A. APPLYING A SAMPLE OF A UROGASTRONE PREPARATION HAVING A SPECIFIC ACITIVITY AS DEFINED AVOVE OF 5-10 MG/KG. TO A COLUMN OF CARBOXYMETHYL CELLULOSE EQUILIBRATED WITH AN 0.01 MOLAR AQUEOUS BUFFER OF PH 4.5, ELUTING THE COLUMN WITH A BUFFER OF PH 5.0 WITH A CONCENTRATION GRADIENT OF 0.01 MOLAR TO 0.5 MOLAR, AND COLLECTING THE ELUTED MATERIAL SHOWING THE BIOLOGICAL ACTIVITY, B. APPLYING THE BIOLOGICAL ACTIVE MATERIAL FROM STEP (A) TO A COLUMN OF A POROUS POLYACRYLAMIDE GEL EXCLUDING MOLECULES OF MOLECULAR WEIGHT GREATER THAN 5,000 IN AN 0.05 MOLAR AQUEOUS BUFFER SOLUTION OF PH 7.2 AND COMPOSED OF VOLATILE COMPONENTS, ELUTING THE COLUMN WITH THE SAME SOLUTION, AND COLLECTING THE ELUTED MATERIAL SHOWING THE BIOLOGICAL ACTIVITY, C. APPLYING THE BIOLOGICALLY ACTIVE MATERIAL FROM STEP (B) TO A COLUMN OF AMINOETHYL CELLULOSE EQUILIBRATED WITH AN 0.01 MOLAR AQUEOUS BUFFER OF PH 5.5 COMPOSED OF VOLATILE COMPONENTS, ELUTING THE COLUMN WITH A BUFFER OF THE SAME MOLARITY AND THEN WITH A BUFFER HAVING A CONCENTRATION GRADIENT OF FROM 0.01 MOLAR TO 0.3 MOLAR, DETECTING THE PRESENCE OF THREE PEAKS OF ELUTED MATERIAL EACH SHOWING THE BIOLOGICAL ACTIVITY AND COLLECTING, FROM THE FIRST ELUTED PEAK MATERIAL WHICH CONTAINS A-UROGASTRONE, FROM THE SECOND ELUTED PEAK MATERIAL WHICH CONTAINS BUROGASTRONE, AND FROM THE THIRD ELUTED PEAK MATERIAL WHICH CONTAINS Y-UROGASTRONE, FOLLOWED BY, IN THE CASE OF MATERIAL FROM THE PEAK FROM STEP (C) CONTAINING BUROGASTRONE, APPLYING THE MATERIAL TO A COLUMN OF CARBOXYMETHYL CELLULOSE EQUILIBRATED WITH AN 0.01 MOLAR AQUEOUS BUFFER OF PH 5.0, ELUTING THE COLUMN WITH A BUFFER OF PH 5.0 WITH A CONCENTRATION GRADIENT OF FROM 0.01 MOLAR TO 0.5 MOLAR, COLLECTING THE ELUTED MATERIAL SHOWING THE BIOLOGICAL ACTIVITY, APPLYING THIS MATERIAL TO A COLUMN OF A POROUS POLYACRYLAMIDE GEL EXCLUDING MOLECULES OF MOLECULAR WEIGHT GREATER THAN 5,000 IN AN 0.05 MOLAR AQUEOUS BUFFER OF PH 7.2 AND COMPOSED OF VOLATILE COMPONENTS, ELUTING THE COLUMN WITH THE SAME SOLUTION AND COLLECTING THE ELUTED MATERIAL SHOWING THE BIOLOGICAL ACTIVITY: OR FOLLOWED BY IN THE CASE OF MATERIAL FROM THE PEAK FROM STEP (C) CONTAINING Y-UROGASTRONE, APPLYING THE MATERIAL TO A COLUMN OF CARBOXYMETHYL CELLULOSE EQUILIBRATED WITH AN 0.01 MOLAR AQUEOUS BUFFER OF PH 4.8, ELUTING THE COLUMN WITH A BUFFER OF PH 4.8 WITH A CONCENTRATION GRADIENT OF FROM 0.01 MOLAR TO 0.5 MOLAR, COLLECTING THE ELUTED MATERIAL SHOWING THE BIOLOGICAL ACTIVITY, APPLYING THIS MATERIAL TO A COLUMN OF A POROUS POLYACRYLAMIDE GEL EXCLUDING MOLECULES OF MOLECULAR WEIGHT GREATER THAN 5,000 IN AN 0.05 MOLAR AQUEOUS BUFFER OF PH 7.2 AND COMPOSED OF VOLATILE COMPONENTS, ELUTING THE COLUMN WITH THE SAME SOLUTION AND COLLECTING THE ELUTED MATERIAL SHOWING THE BIOLOGICAL ACTIVITY.