Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
126 words
A process for large-scale production of an antimicrobial peptide having the sequence Phe-Lys-Cys-Arg-Arg-Trp-Gln-Trp-Arg-Met-Lys-Lys-Leu-Gly-Ala-Pro-Ser-Ile-Thr-Cys-Val-Arg-Arg-Ala-Phe in high purity which comprises the steps of: a) contacting a mixture of peptides containing the antimicrobial peptide which is prepared by enzymatically hydrolysing bovine lactoferrin or bovine milk protein containing lactoferrin with a carboxyl moiety-containing cation-exchange chromatographic medium to adsorb the antimicrobial peptide, b) rinsing the chromatographic medium with a buffer to elute peptides other than the antimicrobial peptide, c) desorbing the antimicrobial peptide in solution from the rinsed cation-exchange chromatographic medium with a solution containing a salt at a concentration of 1 to 4 moles per liter at pH 7-8, and d) desalting the desorbed antimicrobial peptide solution, whereby the antimicrobial peptide of at least 90% by weight purity is obtained.
Claim 2
Dependent Claim
126 words
A process for large-scale production of an antimicrobial peptide having the sequence Phe-Lys-Cys-Arg-Arg-Trp-Gln-Trp-Arg-Met-Lys-Lys-Leu-Gly-Ala-Pro-Ser-Ile-Thr-Cys-Val-Arg-Arg-Ala-Phe in high purity which comprises the steps of: a) contacting a mixture of peptides containing the antimicrobial peptide which is prepared by enzymatically hydrolysing bovine lactoferrin or bovine milk protein containing lactoferrin with a carboxyl moiety-containing cation-exchange chromatographic medium to adsorb the antimicrobial peptide, b) rinsing the chromatographic medium with a buffer to elute peptides other than the antimicrobial peptide, c) desorbing the antimicrobial peptide in solution from the rinsed cation-exchange chromatographic medium with a solution containing a salt at a concentration of 1 to 4 moles per liter at pH 7-8, and d) desalting the desorbed antimicrobial peptide solution, whereby the antimicrobial peptide of at least 90% by weight purity is obtained.
Claim 3
Dependent Claim
75 words
The process according to claim 1, wherein the chromatographic medium is subjected to rinsing with a buffer selected from the group consisting of NaH 2 PO 4 -Na 2 HOP 4 , KH 2 PO 4 -K 2 HPO 4 , NaH 2 PO 4 -K 2 HPO 4 , KH 2 PO 4 -Na 2 HPO 4 , citric acid-Na 2 HPO 4 , acetic acid-sodium acetate and Glycine-NaOH at a pH of 7.8.
Claim 4
Dependent Claim
38 words
The process according to claim 2, wherein the chromatographic medium is subjected to desorbing the antimicrobial peptide with a solution containing a salt selected from the group consisting of ammonium chloride, ammonium acetate, sodium chloride, and potassium chloride.
Claim 5
Dependent Claim
124 words
A process for large-scale production of an antimicrobial peptide having the sequence Phe-Lys-Cys-Arg-Arg-Trp-Gln-Trp-Arg-Met-Lys-Lys-Leu-Gly-Ala-Pro-Ser-Ile-Thr-Cys-Val-Arg-Arg-Ala-Phe in high purity which comprises the steps of: a) contacting a mixture of peptides containing the antimicrobial peptide, which is prepared by enzymatically hydrolysing bovine lactoferrin or bovine milk protein containing lactoferrin with a hydrophobic chromatographic medium, to adsorb the antimicrobial peptide, b) rinsing the chromatographic medium with a buffer at a pH of 6.5-7.5 to elute peptides other than the antimicrobial peptide, c) desorbing the antimicrobial peptide in solution from the rinsed hydrophobic chromatographic medium with a dilute solution of acid or a buffer at pH 4.5-5.5, and d) desalting the desorbed antimicrobial peptide solution, whereby the antimicrobial peptide having the purity of at least 90% by weight is obtained.
Claim 6
Dependent Claim
124 words
A process for large-scale production of an antimicrobial peptide having the sequence Phe-Lys-Cys-Arg-Arg-Trp-Gln-Trp-Arg-Met-Lys-Lys-Leu-Gly-Ala-Pro-Ser-Ile-Thr-Cys-Val-Arg-Arg-Ala-Phe in high purity which comprises the steps of: a) contacting a mixture of peptides containing the antimicrobial peptide, which is prepared by enzymatically hydrolysing bovine lactoferrin or bovine milk protein containing lactoferrin with a hydrophobic chromatographic medium, to adsorb the antimicrobial peptide, b) rinsing the chromatographic medium with a buffer at a pH of 6.5-7.5 to elute peptides other than the antimicrobial peptide, c) desorbing the antimicrobial peptide in solution from the rinsed hydrophobic chromatographic medium with a dilute solution of acid or a buffer at pH 4.5-5.5, and d) desalting the desorbed antimicrobial peptide solution, whereby the antimicrobial peptide having the purity of at least 90% by weight is obtained.
Claim 7
Dependent Claim
36 words
The process according to claim 4, wherein the hydrophobic chromatographic medium contains a moiety selected from the group consisting of a butyl moiety, a hexyl moiety, an octyl moiety, a phenyl moiety, and an octylamine moiety.
Claim 8
Dependent Claim
74 words
The process according to claim 4, wherein the hydrophobic chromatographic medium is subjected to rinsing with a buffer selected from the group consisting of NaH 2 PO 4 -Na 2 HPO 4 , KH 2 PO 4 -K 2 HPO 4 , NaH 2 PO 4 -K 2 HPO 4 , KH 2 PO 4 -Na 2 HPO 4 , citric acid-Na 2 HPO 4 , and acetic acid-sodium acetate at pH of 6.5-7.5.
Claim 9
Dependent Claim
72 words
The process according to claim 4, wherein the antimicrobial peptide is desorbed with a buffer selected from the group consisting of NaH 2 PO 4 -Na 2 HPO 4 , KH 2 PO 4 -K 2 HPO 4 , NaH 2 PO 4 -K 2 HPO 4 , KH 2 PO 4 -Na 2 HPO 4 , citric acid-Na 2 HPO 4 , and acetic acid-sodium acetate at a pH of 4.5-5.5.
Claim 10
Dependent Claim
40 words
The process according to claim 4, wherein the antimicrobial peptide is desorbed with a dilute solution of acid selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, and citric acid at a pH of 4.5-5.5.
Claim 11
Dependent Claim
32 words
The process according to claim 1, wherein the desalting is performed by electrodialysis or ultrafiltration using a semipermeable membrane or hollow fiber cartridge having a molecular weight cut-off of 3000 or less.
Claim 12
Dependent Claim
41 words
The process according to claim 10, wherein the desalting is performed by contacting the desorbed antimicrobial peptide with a hydrophobic chromatographic medium, rinsing the medium with water, and desorbing the antimicrobial peptide from the medium with an aqueous solution of acid.
Claim 13
Dependent Claim
32 words
The process according to claim 4, wherein the desalting is performed by electrodialysis or ultrafiltration using a semipermeable membrane or hollow fiber cartridge having a molecular weight cut-off or 3000 or less.
Claim 14
Dependent Claim
41 words
The process according to claim 4, wherein the desalting is performed by contacting the desorbed antimicrobial peptide with a hydrophobic chromatographic medium, rinsing the medium with water, and desorbing the antimicrobial peptide from the medium with an aqueous solution of acid.