Virus-inactivated HGI-glycoprotein capable of stimulating proliferation and differentiation of human granulocyte, process for preparing same and leukopenia curative containing same

A process for producing a virus-inactivated glycoprotein which stimulates human bone marrow cells to form colonies of granulocytes, which comprises concentrating urine with of normal humans respect to proteins contained therein, contacting the urinary proteins with a cation exchanger to remove impurities by adsorption on said exchanger, contacting the effluent with an anion exchanger to adsorb the active material, eluting the active material with a saline solution according to linear concentration gradient elution, subjecting the eluate to gel filtration chromatography on a highly crosslinked polymer gel to develop the active material collecting fractions of a relative effluent of 1.11 to 1.60, adjusting the aqueous solution containing the collected fractions to pH 5-9, and heating the resulting aqueous solution at 50.degree.-70.degree. C. for 8-30 hours in the presence of albumin of the human origin to inactivate viruses.

A process for producing a virus-inactivated glycoprotein which stimulates human bone marrow cells to form colonies of granulocytes, which comprises concentrating human urine with respect to proteins contained therein, contacting the urinary proteins with a cation exchanger to remove impurities by adsorption on said exchanger, contacting the effluent with an anion exchanger to adsorb the active material, eluting the active material with a saline silution according to linear concentration gradient elution, subjecting the eluate to gel filtration chromatography on a highly crosslinked polymer gel to develop the active material collecting fractions of a relative effluent of 1.11 to 1.60, subjecting the collected fractions to affinity chromatography with a sugar affinitive absorbent to adsorb the effective ingredient, eluting the adsorbed active material with a 20-100 mM saccharide solution, adjusting the aqueous solution containing the collected fractions to pH 5-9, and heating the resulting aqueous solution at 50.degree.-70.degree. C. for 8-30 hours in the presence of albumin of the human origin to inactivate viruses.

A process for producing a virus-inactivated glycoprotein which stimulates human bone marrow cells to form colonies of granulocytes, which comprises concentrating human urine with respect to proteins contained therein, contacting the urinary proteins with a cation exchanger to remove impurities by adsorption on said exchanger, contacting the effluent with an anion exchanger to adsorb the active material, eluting the active material with a saline solution according to linear concentration gradient elution, subjecting the eluate to gel filtration chromatography on a highly crosslinked polymer gel to develop the active material, collecting fractions of a relative effluent of 1.11 to 1.60, subjecting the collected fractions to affinity chromatography with a sugar affinitive absorbent to absorb the active material, eluting the adsorbed active material with a 20-100 mM saccharide solution, subjecting the eluate to preparative zone electrophoresis, eluting the active material with a salt solution, adjusting the eluate to pH 5-9, and heating the resulting eluate at 50.degree.-70.degree. C. for 8-30 hours in the presence of albumin of the human origin to inactivate viruses.

A process for producing a virus-inactivated glycoprotein which stimulates human bone marrow cells to form colonies of granulocytes, which comprises concentrating human urine with respect to proteins contained therein, contacting the urinary proteins with a cation exchanger to remove impurities by adsorption on said exchanger, contacting the effluent with an anion exchanger to adsorb the active material, eluting the active material with a saline solution according to linear concentration gradient elution, subjecting the eluate to gel filtration chromatography on a highly crosslinked polymer gel to develop the active material, collecting fractions of a relative effluent of 1.11 to 1.60, subjecting the collected fractions to affinity chromatography with a sugar affinitive absorbent to absorb the active material, eluting the adsorbed active material with a 20-100 mM saccharide solution, subjecting the eluate to preparative zone electrophoresis, eluating the active material with saline solution, adjusting the concentration of active material in the eluate to at least 70 mg/ml and the pH to 5-9, and heating the resulting eluate at 50.degree.-70.degree. C. for 8-30 hours to inactivate viruses.

A process for producing a virus-inactivated glycoprotein which stimulates human bone marrow cells to form colonies of granulocytes, which comprises concentrating human urine with respect to proteins contained therein, contacting the urinary proteins with a cation exchanger to remove impurities by adsorption on said exchanger, contacting the effluent with an anion exchanger to adsorb the active material, eluting the active material with a saline solution according to linear concentration gradient elution, subjecting the eluate to gel filtration chromatography on a highly crosslinked polymer gel to develop the active material, and collecting fractions of a relative effluent of 1.11 to 1.60;

• the active material containing fraction including the concentrated human urine with respect to urinary proteins in any of the above steps being subjected to virus inactivating treatment by heating the fraction in the form of aqueous solution at 50.degree.-70.degree. C. for 8-30 hours under such conditions that the protein content of the aqueous solution has been adjusted to at least 70 mg/ml and the pH to 5-9.

A process for producing a virus-inactivated glycoprotein which stimulates human bone marrow cells to form colonies of granulocytes, which comprises concentrating human urine with respect to proteins contained therein, contacting the urinary proteins with a cation exchanger to remove impurities by adsorption on said exchanger, contacting the effluent with an anion exchanger to absorb the active material, eluting the active material with a saline solution according to linear concentration gradient elution, subjecting the eluate to gel filtration chromatography on a highly crosslinked polymer gel to develop the active material, collecting fractions of a relative effluent of 1.11 to 1.60, subjecting the collected fractions to affinity chromatography with a sugar affinitive absorbent to adsorb the active material, and eluting the adsorbed active material with a 20-100 mM saccharide solution, the active material containing fraction including the concentrated human urine with respect to urinary protein in any of the above steps being subjected to virus inactivating treatment by heating the fraction in the form of aqueous solution at 50.degree.-70.degree. C. for 8-30 hours under such conditions that the protein content of the aqueous solution has been adjusted to at least 70 mg/ml and the pH 5-9.