# US 4,393,140 A · Process for the preparation of the highly pure enzyme kallikrein from swine pancreas extracts

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- Number: US 4,393,140 A
- Document ID: US4393140A
- Status: Granted
- Kind: A
- Application number: 06/339,017
- Filed: 1982-01-12
- Granted: 1983-07-12
- Earliest claimed: 1981-01-31
- Assignees: Bayer Aktiengesellschaft
- Inventors: Hermann Schutt
- Web page: https://patentgrep.com/patent/US4393140A

## CPC

- C12N 9/6445: {Kallikreins (3.4.21.34; 3.4.21.35)}
- Y10S 435/815: by sorption
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

The invention relates to a process for the preparation of the highly pure enzyme kallikrein from organ extracts, in particular from swine pancreas extracts, the accompanying proteolytic activity of the swine pancreas extract being separated off by chromatography on ion exchangers and the highly pure enzyme kallikrein being obtained by chromatography on support materials with a kallikrein-trypsin inhibitor (basic pancreatic trypsin inhibitor=BPTI) covalently bonded in a particular manner.

## Claims

### Claim 1

A process for the preparation of pure kallikrein, which comprises adjusting an organ extract containing kallikrein to a conductivity of 2 to 6 mS cm.sup.-1 at a pH value of 5 to 8, bringing the organ extract thus prepared into contact with an ion exchanger in batch form or column form, eluting the kallikrein by continuously or discontinuously increasing the ionic strength by means of a mono- or divalent alkali metal salt or alkaline earth metal salt or by lowering the pH value and subsequently purifying the eluate by affinity chromatography on BPTI covalently bonded to a carrier and then desorbing from the BPTI-carrier at a pH value of 4.0 to 5.0.

### Claim 2 (depends on 1)

A process according to claim 1, in which the ion exchanger used is based on cellulose, dextran or agarose and has been crosslinked by treatment with bifunctional reagents.

### Claim 3 (depends on 1, 2)

A process according to claim 1 or 2, in which the carrier to which the BPTI is covalent bonded is based on cellulose, dextran or agarose and has been crosslinked by treatment with bifunctional reagents.

### Claim 4 (depends on 2)

A process according to claim 2, in which the bifunctional reagent is a halohydrin, bis-alkyl halide, bifunctional epoxide or divinyl compound.

### Claim 5 (depends on 3)

A process according to claim 3, in which the bifunctional reagent in halohydrin, bis-alkyl halide, bifunctional epoxide or divinyl compound.