Protein purification process and product

Claim 2

Human leukocyte interferon as a homogenous protein species having a specific activity of from 0.9×10 8 -4×10 8 Units/mg. of protein when assayed on the MDBK bovine cell line and from 2×10 6 -7.6×10 8 Units/mg. of protein when assayed on the AG 1732 human cell line.

Claim 3

The homogeneous human interferon of claim 1 which is a human leukocyte interferon characterized by containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . (II) . . . Tyr-Phe-Gln-Arg . . . (III) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , and (IV) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . .

Claim 4

Human leukocyte interferon as a mixture of human leukocyte interferon species according to claim 1.

Claim 5

A pharmaceutical preparation suitable for parenteral administration for treatment of viral and neoplastic disease states said preparation comprising a minor, effective amount of at least one human leukocyte interferon species according to claim 1 and a major amount of a conventional pharmaceutical parenteral carrier material.

Claim 6

A species of human leukocyte interferon designated α 1 being a homogeneous protein characterized by the following: (a) an amino terminal sequence as follows: Ser-ASp-Leu-Pro-Gln-Thr-His-Ser-Leu-Gly-Asn-Arg-Arg-Thr-Leu-Met-Leu-Leu-Ala-Glu-Met-Arg- . . . (b) a specific activity on MDBK (bovine cells) of about 2.6×10 8 Units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 2.6×10 8 units/mg; (d) a molecular weight of about 16,500±1000 by polyacrylamide gel electrophoresis (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 31% n-propanol (0-40% gradient) in a 1M pyridine--2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) an amino acid composition (±15%) as based on a molecular weight of 16,500 as contained in the following table: Asx: 13.8 Thr: 7.7 Ser: 9.2 Glx: 20.3 Pro: 6.1 Gly: 5.1 Ala: 8.4 Val: 7.4 Met: 3.7 ILeu: 7.4 Leu: 18.0 Tyr: 4.0 Phe: 6.9 His: 3.0 Lys: 10.5 Arg: 6.2 Cys: 3.9 (i) providing peptide fragments upon treatment with trypsin, and fragments eluting as peaks at 3(L), 4(L), 4.2(M), 11.5(M), 12.5(S), 14.5(M), 16(S), 18(M), 20(S), 21(S), 22.5(S) and 29(M)% n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (j) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 7

A species of human leukocyte interferon designated α 1 being a homogeneous protein characterized by the following: (a) an amino terminal sequence as follows: Ser-ASp-Leu-Pro-Gln-Thr-His-Ser-Leu-Gly-Asn-Arg-Arg-Thr-Leu-Met-Leu-Leu-Ala-Glu-Met-Arg- . . . (b) a specific activity on MDBK (bovine cells) of about 2.6×10 8 Units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 2.6×10 8 units/mg; (d) a molecular weight of about 16,500±1000 by polyacrylamide gel electrophoresis (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 31% n-propanol (0-40% gradient) in a 1M pyridine--2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) an amino acid composition (±15%) as based on a molecular weight of 16,500 as contained in the following table: Asx: 13.8 Thr: 7.7 Ser: 9.2 Glx: 20.3 Pro: 6.1 Gly: 5.1 Ala: 8.4 Val: 7.4 Met: 3.7 ILeu: 7.4 Leu: 18.0 Tyr: 4.0 Phe: 6.9 His: 3.0 Lys: 10.5 Arg: 6.2 Cys: 3.9 (i) providing peptide fragments upon treatment with trypsin, and fragments eluting as peaks at 3(L), 4(L), 4.2(M), 11.5(M), 12.5(S), 14.5(M), 16(S), 18(M), 20(S), 21(S), 22.5(S) and 29(M)% n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (j) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 8

A species of human leukocyte interferon designated α 2 being a homogeneous protein characterized by the following: (a) a blocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 4.0×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 3×10 8 units/mg; (d) a molecular weight of about 16,200±1000 by polyacrylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 32% n-propanol (0-40% gradient) in a 1M pyridine--2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) resistant to deactivation by treatment with leucine aminopeptidase and aminopeptidase M; (i) an amino acid composition (±15%) as based on a molecular weight of 16,200 as contained in the following table: Asx: 11.7 Thr: 8.3 Ser: 10.0 Glx: 20.5 Pro: 4.9 Gly: 4.5 Ala: 8.3 Val: 6.2 Met: 3.8 ILeu: 7.0 Leu: 18.0 Tyr: 4.4 Phe: 8.0 His: 2.8 Lys: 9.0 Arg: 7.1 Cys: 4.0 (j) providing peptide fragments upon treatment with trypsin said fragments eluted as peaks at 3(L), 4(L), 4.2(M), 11.5(M), 12.5(S), 14.5(M), 16(S), 18(M), 27(S) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; (k) eluted as a single peak at a concentration of 68% n-propanol (72.5-50% gradient) in 0.1M sodium acetate aqueous buffer from a glycerol bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.25 ml/min. and (l) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 9

A species of human leukocyte interferon designated α 2 being a homogeneous protein characterized by the following: (a) a blocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 4.0×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 3×10 8 units/mg; (d) a molecular weight of about 16,200±1000 by polyacrylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 32% n-propanol (0-40% gradient) in a 1M pyridine--2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) resistant to deactivation by treatment with leucine aminopeptidase and aminopeptidase M; (i) an amino acid composition (±15%) as based on a molecular weight of 16,200 as contained in the following table: Asx: 11.7 Thr: 8.3 Ser: 10.0 Glx: 20.5 Pro: 4.9 Gly: 4.5 Ala: 8.3 Val: 6.2 Met: 3.8 ILeu: 7.0 Leu: 18.0 Tyr: 4.4 Phe: 8.0 His: 2.8 Lys: 9.0 Arg: 7.1 Cys: 4.0 (j) providing peptide fragments upon treatment with trypsin said fragments eluted as peaks at 3(L), 4(L), 4.2(M), 11.5(M), 12.5(S), 14.5(M), 16(S), 18(M), 27(S) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; (k) eluted as a single peak at a concentration of 68% n-propanol (72.5-50% gradient) in 0.1M sodium acetate aqueous buffer from a glycerol bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.25 ml/min. and (l) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 10

A species of human leukocyte interferon designated β 1 being a homogeneous protein characterized by the following: (a) a blocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 1.7×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 2.2×10 8 units/mg; (d) a molecular weight of about 16,000(±1000) by polyacrylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 31% n-propanol (0-40% gradient) in a 1M pyridine--2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min.; (h) resistant to deactivation by treatment with leucine aminopeptidase and aminopeptidase M; (i) an amino acid composition (±15%) as based on a molecular weight of 16,000 as contained in the following table: Asx: 11.3 Thr: 8.7 Ser: 10.3 Glx: 20.6 Pro: 5.2 Gly: 4.9 Ala: 7.2 Val: 6.0 Met: 4.8 Ile: 6.4 Leu: 18.1 Tyr: 4.4 Phe: 8.6 His: 2.8 Lys: 8.7 Arg: 7.9 Cys: 3.0 (j) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 11.5(M), 12.5(S), 14.5(M), 16(S), 17.5(S), 18(M) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (k) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (IV) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His Lys) . . . .

Claim 11

A species of human leukocyte interferon designated β 1 being a homogeneous protein characterized by the following: (a) a blocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 1.7×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 2.2×10 8 units/mg; (d) a molecular weight of about 16,000(±1000) by polyacrylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 31% n-propanol (0-40% gradient) in a 1M pyridine--2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min.; (h) resistant to deactivation by treatment with leucine aminopeptidase and aminopeptidase M; (i) an amino acid composition (±15%) as based on a molecular weight of 16,000 as contained in the following table: Asx: 11.3 Thr: 8.7 Ser: 10.3 Glx: 20.6 Pro: 5.2 Gly: 4.9 Ala: 7.2 Val: 6.0 Met: 4.8 Ile: 6.4 Leu: 18.1 Tyr: 4.4 Phe: 8.6 His: 2.8 Lys: 8.7 Arg: 7.9 Cys: 3.0 (j) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 11.5(M), 12.5(S), 14.5(M), 16(S), 17.5(S), 18(M) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (k) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (IV) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His Lys) . . . .

Claim 12

A species of human leukocyte interferon designated β 2 being a homogeneous protein characterized by the following: (a) an unblocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 4.0×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 2×10 8 units/mg; (d) a molecular weight of about 16,500 (±1000) by polyacylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 32% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) an amino acid composition (±15%) as based on molecular weight of 16,500 as contained in the following table: Asx: 11.5 Thr: 9.0 Ser: 10.0 Glx: 21.9 Pro: 5.0 Gly: 5.0 Ala: 7.9 Val: 6.7 Met: 4.4 ILeu: 7.4 Leu: 18.9 Tyr: 4.6 Phe: 8.7 His: 3.0 Lys: 8.5 Arg: 8.0 Cys: 1.8 (i) providing peptide fragments upon treatment with trypsin, said fragments eluting at peaks at 3(L), 4(L), 4.2(M), 11.5(M), 12.5(S), 14.5(M), 16(S), 17.5(S), 18(M) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; (j) eluted as a single peak at a concentration of 66.5% n-propanol (72.5-50% gradient) in 0.1M sodium acetate aqueous buffer from a glycerol bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.25 ml/min.; and (k) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 13

A species of human leukocyte interferon designated β 2 being a homogeneous protein characterized by the following: (a) an unblocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 4.0×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 2×10 8 units/mg; (d) a molecular weight of about 16,500 (±1000) by polyacylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 32% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) an amino acid composition (±15%) as based on molecular weight of 16,500 as contained in the following table: Asx: 11.5 Thr: 9.0 Ser: 10.0 Glx: 21.9 Pro: 5.0 Gly: 5.0 Ala: 7.9 Val: 6.7 Met: 4.4 ILeu: 7.4 Leu: 18.9 Tyr: 4.6 Phe: 8.7 His: 3.0 Lys: 8.5 Arg: 8.0 Cys: 1.8 (i) providing peptide fragments upon treatment with trypsin, said fragments eluting at peaks at 3(L), 4(L), 4.2(M), 11.5(M), 12.5(S), 14.5(M), 16(S), 17.5(S), 18(M) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; (j) eluted as a single peak at a concentration of 66.5% n-propanol (72.5-50% gradient) in 0.1M sodium acetate aqueous buffer from a glycerol bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.25 ml/min.; and (k) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 14

A species of human leukocyte interferon designated β 3 being a homogeneous protein characterized by the following: (a) a specific activity on MDBK (bovine cells) of about 3.6×10 8 units/mg; (b) a specific activity on AG1732 (human line) cells of about 7.6×10 8 units/mg; (c) having a major molecular weight band of about 24,000 (±1000) by polyacrylamide gel electrophoresis; (d) an amino sugar content of less than 1 residue per molecule; (e) positive growth inhibition activity; (f) eluted as a single peak at a concentration of 32% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (g) an amino acid composition as contained in the following table: Asx: 18.7 Thr: 8.3 Ser: 16.9 Glx: 35.0 Pro: 6.5 Gly: 3.5 Ala: 11.5 Val: 8.3 Met: 6.8 ILeu: 11.6 Leu: 26.0 Tyr: 6.7 Phe: 12.0 His: 4.3 Lys: 13.2 Arg: 12.1 Cys: - (h) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 4.2(M), 4.5(S), 10(M), 12.5(S), 14(S), 14.5(M), 16(S), 18(L), 19.5(M), 27(M) and 32(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (i) containing the following amino sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (IV) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . .

Claim 15

A species of human leukocyte interferon designated β 3 being a homogeneous protein characterized by the following: (a) a specific activity on MDBK (bovine cells) of about 3.6×10 8 units/mg; (b) a specific activity on AG1732 (human line) cells of about 7.6×10 8 units/mg; (c) having a major molecular weight band of about 24,000 (±1000) by polyacrylamide gel electrophoresis; (d) an amino sugar content of less than 1 residue per molecule; (e) positive growth inhibition activity; (f) eluted as a single peak at a concentration of 32% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (g) an amino acid composition as contained in the following table: Asx: 18.7 Thr: 8.3 Ser: 16.9 Glx: 35.0 Pro: 6.5 Gly: 3.5 Ala: 11.5 Val: 8.3 Met: 6.8 ILeu: 11.6 Leu: 26.0 Tyr: 6.7 Phe: 12.0 His: 4.3 Lys: 13.2 Arg: 12.1 Cys: - (h) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 4.2(M), 4.5(S), 10(M), 12.5(S), 14(S), 14.5(M), 16(S), 18(L), 19.5(M), 27(M) and 32(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (i) containing the following amino sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (IV) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . .

Claim 16

A species of human leukocyte interferon designated γ 1 being a homogeneous protein characterized by the following: (a) a blocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 2.6×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 2×10 8 units/mg; (d) a molecular weight of about 17,700±1000 by polyacrylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 31% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) resistant to deactivation by treatment with leucine aminopeptidase and aminopeptidase M; (i) an amino acid composition (±15%) as based on a molecular weight of 17,700 contained in the following table: Asx: 13.1 Thr: 8.4 Ser: 10.2 Glx: 23.9 Pro: 4.5 Gly: 5.7 Ala: 8.7 Val: 7.3 Met: 4.3 ILeu: 7.9 Leu: 20.3 Tyr: 4.8 Phe: 8.6 His: 3.7 Lys: 10.1 Arg: 8.0 Cys: 3.3 (j) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 4.2(M), 4.5(S), 5(S), 6.5(S), 11.5(S), 12.5(S), 14.5(M), 16(S), 17.5(M), 18(M) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (k) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 17

A species of human leukocyte interferon designated γ 1 being a homogeneous protein characterized by the following: (a) a blocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 2.6×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 2×10 8 units/mg; (d) a molecular weight of about 17,700±1000 by polyacrylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 31% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) resistant to deactivation by treatment with leucine aminopeptidase and aminopeptidase M; (i) an amino acid composition (±15%) as based on a molecular weight of 17,700 contained in the following table: Asx: 13.1 Thr: 8.4 Ser: 10.2 Glx: 23.9 Pro: 4.5 Gly: 5.7 Ala: 8.7 Val: 7.3 Met: 4.3 ILeu: 7.9 Leu: 20.3 Tyr: 4.8 Phe: 8.6 His: 3.7 Lys: 10.1 Arg: 8.0 Cys: 3.3 (j) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 4.2(M), 4.5(S), 5(S), 6.5(S), 11.5(S), 12.5(S), 14.5(M), 16(S), 17.5(M), 18(M) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (k) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys) . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 18

A species of human leukocyte interferon designated γ 2 being a homogeneous protein characterized by the following: (a) a blocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 4.0×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 1.5×10 8 units/mg; (d) a molecular weight of about 17,700 (±1000) on polyacrylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 32% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) resistant to deactivation by treatment with leucine aminopeptidase and aminopeptidase M; (i) an amino acid composition (±15%) as based on a molecular weight of about 17,700 as contained in the following table: Asx: 13.3 Thr: 9.6 Ser: 7.8 Glx: 25.0 Pro: 4.8 Gly: 5.0 Ala: 8.1 Val: 7.0 Met: 3.9 ILeu: 8.0 Leu: 20.1 Tyr: 4.8 Phe: 9.0 His: 3.3 Lys: 10.0 Arg: 8.5 Cys: 2.9 (j) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 4.2(M), 4.5(S), 5(S), 11.5(S), 12.5(S), 14.5(S), 16(S), 18(L) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (k) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys . . . .

Claim 19

A species of human leukocyte interferon designated γ 2 being a homogeneous protein characterized by the following: (a) a blocked amino terminus; (b) a specific activity on MDBK (bovine cells) of about 4.0×10 8 units/mg; (c) a specific activity on Ag 1732 (human line) cells of about 1.5×10 8 units/mg; (d) a molecular weight of about 17,700 (±1000) on polyacrylamide gel electrophoresis; (e) an amino sugar content of less than 1 residue per molecule; (f) positive growth inhibition activity; (g) eluted as a single peak at a concentration of 32% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (h) resistant to deactivation by treatment with leucine aminopeptidase and aminopeptidase M; (i) an amino acid composition (±15%) as based on a molecular weight of about 17,700 as contained in the following table: Asx: 13.3 Thr: 9.6 Ser: 7.8 Glx: 25.0 Pro: 4.8 Gly: 5.0 Ala: 8.1 Val: 7.0 Met: 3.9 ILeu: 8.0 Leu: 20.1 Tyr: 4.8 Phe: 9.0 His: 3.3 Lys: 10.0 Arg: 8.5 Cys: 2.9 (j) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 4.2(M), 4.5(S), 5(S), 11.5(S), 12.5(S), 14.5(S), 16(S), 18(L) and 29(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (k) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Lys-Tyr-Phe-Gln-Arg . . . , (III) . . . Glx-Asx-Ser-Ile-Leu-Ala-Val-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VI) . . . (Ser-2, Ile, Leu-2, Phe, Lys, Cys) . . . , (VII) . . . (Asx-2, Thr-2, Ser, Glx-3, Pro, Gly, Ala, Val, Leu-3, Tyr, Lys . . . .

Claim 20

A species of human leukocyte interferon designated γ 3 being a homogeneous protein characterized by the following: (a) a specific activity on MDBK (bovine cells) of about 3.5×10 8 units/mg.; (b) a specific activity on AG1732 (human line) cells of about 1.5×10 7 units/mg; (c) haing a major molecular weight band of about 17,200 (±1000) on polyacrylaminde gel electrophoresis; (d) an amino sugar content of less than 1 residue per molecule; (e) positive growth inhibition activity; (f) eluted as a single peak at a concentration of 34% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 mg/min; (g) an amino acid composition as contained in the following table; Asx: 15.0 Thr: 8.6 Ser: 10.1 Glx: 22.8 Pro: 4.8 Gly: 3.2 Ala: 9.3 Val: 5.5 Met: 5.6 ILeu: 6.8 Leu: 20.5 Tyr: 3.7 Phe: 7.2 His: 2.9 Lys: 7.6 Arg: 10.1 Cys: 3.1 (h) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(M), 4(M), 4.2(M), 11.5(M), 12.5(S), 13.5(S), 14.5(M), 16(S), 18(L), 20(S) and 32(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (i) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Tyr-Phe-Gln-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 21

A species of human leukocyte interferon designated γ 3 being a homogeneous protein characterized by the following: (a) a specific activity on MDBK (bovine cells) of about 3.5×10 8 units/mg.; (b) a specific activity on AG1732 (human line) cells of about 1.5×10 7 units/mg; (c) haing a major molecular weight band of about 17,200 (±1000) on polyacrylaminde gel electrophoresis; (d) an amino sugar content of less than 1 residue per molecule; (e) positive growth inhibition activity; (f) eluted as a single peak at a concentration of 34% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 mg/min; (g) an amino acid composition as contained in the following table; Asx: 15.0 Thr: 8.6 Ser: 10.1 Glx: 22.8 Pro: 4.8 Gly: 3.2 Ala: 9.3 Val: 5.5 Met: 5.6 ILeu: 6.8 Leu: 20.5 Tyr: 3.7 Phe: 7.2 His: 2.9 Lys: 7.6 Arg: 10.1 Cys: 3.1 (h) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(M), 4(M), 4.2(M), 11.5(M), 12.5(S), 13.5(S), 14.5(M), 16(S), 18(L), 20(S) and 32(M) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient; and (i) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Tyr-Phe-Gln-Arg . . . , (IV) . . . Ile-Thr-Leu-Tyr-Leu-Lys . . . , (V) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . , (VIII) . . . (Asx, Thr-2, Ser, Glx-3, Pro, Ala, Val, Met, Ile-2, Leu-2, Phe-2, His, Lys) . . . .

Claim 22

A species of human leukocyte interferon designated γ 4 being a homogeneous protein characterized by the following: (a) a specific activity on MDBK (bovine cells) of about 3.5×10 8 units/mg; (b) a specific activity on AG1732 (human line) cells of about 4.0×10 8 units/mg; (c) a molecular weight of about 21,000 (±1000) on polyacrylamide gel electrophoresis; (d) an amino sugar content of less than 1 residue per molecule; (e) positive growth inhibition activity; (f) eluted as a single peak at a concentration of 35% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from a octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (g) an amino acid composition as contained in the following table: Asx: 17.9 Thr: 7.3 Ser: 13.5 Glx: 27.0 Pro: 6.5 Gly: 4.8 Ala: 10.4 Val: 7.9 Met: 4.9 ILeu: 9.7 Leu: 24.1 Tyr: 5.0 Phe: 9.1 His: 3.8 Lys: 12.3 Arg: 8.5 Cys: 4.1 and (h) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Tyr-Phe-Gln-Arg . . . , (III) . . . Ile, Thr-Leu-Tyr-Leu-Lys . . . , (IV) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . .

Claim 23

A species of human leukocyte interferon designated γ 4 being a homogeneous protein characterized by the following: (a) a specific activity on MDBK (bovine cells) of about 3.5×10 8 units/mg; (b) a specific activity on AG1732 (human line) cells of about 4.0×10 8 units/mg; (c) a molecular weight of about 21,000 (±1000) on polyacrylamide gel electrophoresis; (d) an amino sugar content of less than 1 residue per molecule; (e) positive growth inhibition activity; (f) eluted as a single peak at a concentration of 35% n-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from a octyl bonded silica matrix HPLC 4.6×250 mm column at room temperature and a flow rate of 0.2 ml/min; (g) an amino acid composition as contained in the following table: Asx: 17.9 Thr: 7.3 Ser: 13.5 Glx: 27.0 Pro: 6.5 Gly: 4.8 Ala: 10.4 Val: 7.9 Met: 4.9 ILeu: 9.7 Leu: 24.1 Tyr: 5.0 Phe: 9.1 His: 3.8 Lys: 12.3 Arg: 8.5 Cys: 4.1 and (h) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Tyr-Phe-Gln-Arg . . . , (III) . . . Ile, Thr-Leu-Tyr-Leu-Lys . . . , (IV) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . .

Claim 24

A species of human leukocyte interferon designated γ 5 being a homogenous protein characterized by the following: (a) a specific activity on MDBK (bovine cells) of about 0.9×10 8 units/mg; (b) a specific activity on AG1732 (human line) cells of about 2×10 6 units/mg; (c) having a major molecular weight band of about 16,500 (±1000) on polyacrylamide gel electrophoresis; (d) an amino sugar content of less than 1 residue per molecule; (e) positive growth inhibition activity; (f) eluted as a single peak at a concentration of 35.5% no-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.5×250 mm column at room temperature and a flow rate of 0.2 ml/min; (g) an amino acid composition as contained in the following table: Asx: 13.8 Thr: 7.6 Ser: 9.0 Glx: 20.8 Pro: 4.2 Gly: 4.0 Ala: 9.3 Val: 5.1 Met: 4.7 ILeu: 6.6 Leu: 19.6 Tyr: 3.6 Phe: 6.4 His: 2.8 Lys: 12.0 Arg: 9.0 Cys: 2.3 (h) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 4.2(M), 4.5(M), 7(S), 7.5(S), 10(S), 11.5(L), 12.5(S), 14(S), 14.5(M), 16(S), 18(L), 24.5(S), 25.5(S) and 32(S) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient. and (i) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Tyr-Phe-Gln-Arg . . . , (III) . . . Ile, Thr-Leu-Tyr-Leu-Lys . . . , (IV) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . .

Claim 25

A species of human leukocyte interferon designated γ 5 being a homogenous protein characterized by the following: (a) a specific activity on MDBK (bovine cells) of about 0.9×10 8 units/mg; (b) a specific activity on AG1732 (human line) cells of about 2×10 6 units/mg; (c) having a major molecular weight band of about 16,500 (±1000) on polyacrylamide gel electrophoresis; (d) an amino sugar content of less than 1 residue per molecule; (e) positive growth inhibition activity; (f) eluted as a single peak at a concentration of 35.5% no-propanol (0-40% gradient) in a 1M pyridine-2M formic acid aqueous buffer from an octyl bonded silica matrix HPLC 4.5×250 mm column at room temperature and a flow rate of 0.2 ml/min; (g) an amino acid composition as contained in the following table: Asx: 13.8 Thr: 7.6 Ser: 9.0 Glx: 20.8 Pro: 4.2 Gly: 4.0 Ala: 9.3 Val: 5.1 Met: 4.7 ILeu: 6.6 Leu: 19.6 Tyr: 3.6 Phe: 6.4 His: 2.8 Lys: 12.0 Arg: 9.0 Cys: 2.3 (h) providing peptide fragments upon treatment with trypsin, said fragments eluting as peaks at 3(L), 4(L), 4.2(M), 4.5(M), 7(S), 7.5(S), 10(S), 11.5(L), 12.5(S), 14(S), 14.5(M), 16(S), 18(L), 24.5(S), 25.5(S) and 32(S) % n-propanol when the reaction media is passed through an octyl bonded silica matrix 10μ particle size HPLC 4.6×250 mm column at room temperature using a 0.03M pyridine-0.1M formic acid aqueous buffer (pH 3) at a flow rate of 0.5 ml/min. with a 1 hour 0 to 40% n-propanol gradient. and (i) containing the following amino acid sequences: (I) . . . Ala-Glu-Ile-Met-Arg . . . , (II) . . . Tyr-Phe-Gln-Arg . . . , (III) . . . Ile, Thr-Leu-Tyr-Leu-Lys . . . , (IV) . . . Thr-Leu-Met-Leu-Leu-Ala-Gln-Met-Arg . . . .