IL46587A - Process for the isolation of a polyvalent proteinase inhibitor - Google Patents
Process for the isolation of a polyvalent proteinase inhibitorInfo
- Publication number
- IL46587A IL46587A IL46587A IL4658775A IL46587A IL 46587 A IL46587 A IL 46587A IL 46587 A IL46587 A IL 46587A IL 4658775 A IL4658775 A IL 4658775A IL 46587 A IL46587 A IL 46587A
- Authority
- IL
- Israel
- Prior art keywords
- proteinase inhibitor
- sulfonic acid
- inhibitor
- adsorbant
- polystyrene
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims description 27
- 239000000137 peptide hydrolase inhibitor Substances 0.000 title claims description 21
- 238000002955 isolation Methods 0.000 title claims description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 17
- 239000003112 inhibitor Substances 0.000 claims description 15
- 229920001577 copolymer Polymers 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 238000010828 elution Methods 0.000 claims description 11
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 10
- 239000011347 resin Substances 0.000 claims description 10
- 229920005989 resin Polymers 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 10
- 150000002500 ions Chemical class 0.000 claims description 9
- 239000011780 sodium chloride Substances 0.000 claims description 9
- 229940117913 acrylamide Drugs 0.000 claims description 8
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 7
- 238000001179 sorption measurement Methods 0.000 claims description 7
- NLVXSWCKKBEXTG-UHFFFAOYSA-M ethenesulfonate Chemical compound [O-]S(=O)(=O)C=C NLVXSWCKKBEXTG-UHFFFAOYSA-M 0.000 claims description 6
- 210000001557 animal structure Anatomy 0.000 claims description 4
- ABLZXFCXXLZCGV-UHFFFAOYSA-N Phosphorous acid Chemical group OP(O)=O ABLZXFCXXLZCGV-UHFFFAOYSA-N 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 229920001467 poly(styrenesulfonates) Polymers 0.000 claims description 3
- 229940005642 polystyrene sulfonic acid Drugs 0.000 claims description 3
- NLVXSWCKKBEXTG-UHFFFAOYSA-N vinylsulfonic acid Chemical compound OS(=O)(=O)C=C NLVXSWCKKBEXTG-UHFFFAOYSA-N 0.000 claims description 3
- 238000004132 cross linking Methods 0.000 claims description 2
- 239000004793 Polystyrene Substances 0.000 claims 2
- 229920002223 polystyrene Polymers 0.000 claims 2
- -1 sulfonic acid ion Chemical class 0.000 claims 2
- 238000007334 copolymerization reaction Methods 0.000 claims 1
- 239000012266 salt solution Substances 0.000 claims 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 claims 1
- 239000002253 acid Substances 0.000 description 12
- 239000000284 extract Substances 0.000 description 7
- 239000001768 carboxy methyl cellulose Substances 0.000 description 6
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 6
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 6
- 229940105329 carboxymethylcellulose Drugs 0.000 description 6
- 229940012957 plasmin Drugs 0.000 description 6
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 5
- 210000000056 organ Anatomy 0.000 description 5
- 239000000047 product Substances 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 210000004072 lung Anatomy 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 238000005227 gel permeation chromatography Methods 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 235000019833 protease Nutrition 0.000 description 2
- HRZFUMHJMZEROT-UHFFFAOYSA-L sodium disulfite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])(=O)=O HRZFUMHJMZEROT-UHFFFAOYSA-L 0.000 description 2
- 235000010262 sodium metabisulphite Nutrition 0.000 description 2
- 125000000542 sulfonic acid group Chemical group 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 1
- 108090000317 Chymotrypsin Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 108060005987 Kallikrein Proteins 0.000 description 1
- 102000001399 Kallikrein Human genes 0.000 description 1
- UEZVMMHDMIWARA-UHFFFAOYSA-N Metaphosphoric acid Chemical compound OP(=O)=O UEZVMMHDMIWARA-UHFFFAOYSA-N 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-N Propionic acid Substances CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 1
- 101710102734 Proteinase inhibitor A Proteins 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 229920006322 acrylamide copolymer Polymers 0.000 description 1
- 230000000274 adsorptive effect Effects 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 239000006286 aqueous extract Substances 0.000 description 1
- 230000003190 augmentative effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 229960002376 chymotrypsin Drugs 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000008098 formaldehyde solution Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- RPACBEVZENYWOL-XFULWGLBSA-M sodium;(2r)-2-[6-(4-chlorophenoxy)hexyl]oxirane-2-carboxylate Chemical compound [Na+].C=1C=C(Cl)C=CC=1OCCCCCC[C@]1(C(=O)[O-])CO1 RPACBEVZENYWOL-XFULWGLBSA-M 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- NBOMNTLFRHMDEZ-UHFFFAOYSA-N thiosalicylic acid Chemical compound OC(=O)C1=CC=CC=C1S NBOMNTLFRHMDEZ-UHFFFAOYSA-N 0.000 description 1
- 229940103494 thiosalicylic acid Drugs 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
- C07K14/8114—Kunitz type inhibitors
- C07K14/8117—Bovine/basic pancreatic trypsin inhibitor (BPTI, aprotinin)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Description
PROCESS FOR THE ISOLATION OF A POLYVALENT PROTEINASE INHIBITOR (HOE 74/B 002) The present invention relates to the isolation of a polyvalent inhibitor of proteinases from animal organs.
The basic polyvalent proteinase inhibitor isolated from ani^ mal organs has an outstanding importance in therapy owing to its property of inhibiting trypsin, chymotrypsin, the kallikreins of plasma, organs and urine, as well as plasmin.
Since the discovery of this inhibitor, a great number of processes for isolating it have become known. After extraction of the organs, the inhibitor is isolated from the extract by fractionation with organic solvents such as alcohol or acetone, with the aid of various protein precipitating agents such as sul-foealicylic acid, thiosalicylic acid, trichloroacetic acid, meta-phosphoric acid or various salts, by specific adsorption or by chromatographic methods. However, most of these processes are unsuitable for an operation on an industrial scale, if it is required to obtain goad yields and to have to p«srform the process in simple way at the same time.
A process which essentially satisfies these two requirements comprises the adsorption of the proteinase inhibitor of the organ extract on carboxymethyl-cellulose. However, the process step of adsorption on carboxymethyl-cellulose requires a low ion strength of the solution which can be attained only by strong dilution of the organ extract. In addition to this disadvantage of having to operate with large volumes and the great amount of ion exchanger, the filtrability of the strongly swelling carboxymethyl-cellulose represents technically a difficult problem which is the more serious in the regeneration which must be carried out in the alkaline pH-range. In view of the known basic properties of the polyvalent proteinase inhibitor, it seemed self-suggesting to use for the isolation of this substance, instead of carboxy-methyl-cellulose, other acid ion exchangers which have the form^ of a resin and which do not have the above-described disadvantage. The methods described in the literature in this respect, however, proved very unsatisfactory. For the adsorption of the inhibitor, a large amount of exchanger resin is required, while the yields are relatively poor and the product obtained has no satisfactory purity, or a very pure product is prepared with considerably expensive fractionation measures without regard to the yield, which would be inapplicable on an industrial scale.
Now, we have found that cation exchangers which carry sulfonic acid groups or phosphonic acid groups as functional groups are particularly suited for the isolation of the polyvalent proteinase inhibitor.
A copolymer gel of ethene-sulfonate and acrylamide may be used with advantage as adsorbant. This gel, which can be prepared according to known processes as described in Example 1 of the present specification, is a copolymer of ethene-sulfonic acid and acrylamide (in a weight proportion of about 20:1 to 2:1, preferably 8:1 to 3:1) which is cross-linked with 1 to 15 % of formaldehyde. The grain size suitable for the adsorption of the proteinase inhibitor is O.06 to 3 mm.
Well suitable for the process of the invention have also proved the commercial polyphenol- (ΛΓ -sulfonic acid ion exchangers, furthermore polystyrene-sulfonic acid resins, in particular those whose capacity is increased owing to a macro—porous structure, or polystyrene-phosphon c acid resins. A grain size of these ion exchangers of about 0.03 to 0.08 mm has shown to be particularly advantageous.
The exchanger types of the present invention are distinguished over carboxymethyl-cellulose by a considerably higher adsorptive capacity. ^ Furthermore, the filtration speed as compared to that with carboxymethyl-cellulose for the separation of not-adsorbed material as well as for the elution of the inhibitor from the resins used according to the invention can be essentially augmented. The ion exchangers mentioned can be easily regenerated after elution of the inhibitor, since the through-flow speed of solutions is also not impeded in the alkaline range. The exchangers may be used several times for the isolation of the polyvalent proteinase inhibitor. This can be effected according to the batch method or according to the column method.
Accordingly, the object of the present invention is a pro- s cess for the isolation of a polyvalent proteinase inhibitor from its aqueous solutions, preferably from an aqueous extract of animal organs, preferably from bovine lungs, which comprises adsorbing the inhibitor on an ion exchanger resin with functional sulfonic acid groups or phosphonic acid groups, at pH-values of from 0.5 to 10.5, preferably pH-values of from 7.5 to 9.5, separating the adsorbant and subsequently eluting it with aqueous saline solutions at pH-values of 1 to 13, preferably pH-values of from 10 to 12.5; in proximity of pH 12.5, the addition of salt may be omitted, whereas with falling pH-value increasing amounts of salt are to be added in order to augment the elutive capacity. If the ethene-sulfonate/acrylamide copolymer gel is used, there are added, for example at a pH-value of 3, about 10% of sodium chloride to the eluting medium. If polyphenol- ^-sulfonic acid ion exchangers or polystyrene-phosphonic acid resins or polystyrene-sulfonic acid acid resins are used, the elution may be carried out, for example at pH 7.0, with addition of about 20% of sodium chloride or such-amounts of another salt which provide a comparable elution ca- ^ pacity. With these types of exchangers, it must be taken into consideration that, owing to the higher salt concentrations required for the elution, precipitations of the inhibitor may occur at elutions at pH-values below 5, which must be compensated by an increased elution volume in order to keep losses of yield as low as possible.
The process of the invention is used in the isolation of the proteinase inhibitor; it is of advantage to use as starting material for the adsorption of the proteinase inhibitor a pre-purified extract of animal organs.
For determining the proteinase inhibitor isolated according to the invention, the inhibition of plasmin with casein as the substrate may be used. This test determines how much the enzy-matical activity of a plasmin, previously determined according to the method described by L.F. Remmert and P.P- Cohen, J. Biol. Chem. 181, 431 (1949), is reduced by a quantity of inhibitor added. The inhibition of a plasmin unit is defined as anti-plasmin unit. With a view to the specific activity of the proteinase inhibitor, the anti-plasmin units (APU)/ml are referred to an extinction value of the solution of the inhibitor of 1.0 measured at a wave length of 280 nm. Consequently, referred to APU/mg of nitrogen, a purification factor of 20 to 35 is attained according to the process of the invention.
The eluate obtained according to the invention may be further purified by gel chromatography, whereupon the inhibitor is obtained in high purity and with good yield. Thus, an inhibitor of about 170 APU/ml at E2so β 1,0 can De obtained« This purity of the polyvalent proteinase inhibitor permits its therapeutic use in humans. t The following Examples illustrate the invention.
EXAMPLE 1: A. Preparation of a copolymer of ethene-sulfonate and acryl amide : 85 parts by weight of 25% aqueous solution of the Na-salt of ethene-sulfonic acid were adjusted to pH 6.9 with 40% H2S04. Then, 5.3 parts by weight of acryl-amide were dissolved in this solution and the whole was heated to 45° C. Polymerization was initiated by the addition of 0.425 part by weight of ammonium-peroxyde-disulfate and 0.085 part by weight of sodium disulfite. The batch heated up to 98° C. In order to complete the polymerization, 0.0425 part by weight of ammonium-peroxyde-disulfate and 0.0085 part by weight of sodium disulfite were added, the whole was stirred for 2 hours at 85° C and then cooled.
B. Cross-linking with formaldehyde and thermical after-treatment 13.6 parts by weight of a 28% formaldehyde solution were added to the solution of copolymer and allowed to stand for 15 hours at 25° C. The gel obtained was dried at 80° C under reduced pressure and the dry product was heated for 30 minutes to 150° C. It was then ground and sieved. The fraction having a grain size of 0,06 to 0.3 mm was found to be especially suitable for the isolation of the polyvalent proteinase inhibitor. Before use, the cross-linked product was swelled by washing with desalted water and freed from water- soluble by-products.
EXAMPLE 2: 33 kg of the wet ethene-sulfonate/acryl-amide copolymer ge^L (prepared as described in Example 1) were added to 1650 1 of a g crude extract of lung tissue which contained 12 x 10 antiplasmin units. The mixture was stirred for 30 minutes and then allowed to stand for 30 minutes to permit sedimentation of the adsorbant. Then, 1250 1 of the supernatant could be decanted off. In order to remove the residual supernatant, the adsorbant was filtered off. Elution of the proteinase inhibitor was effected with 120 1 of de- ionized water, the pH-value of the suspension having been adjusted to 12.5 by means of concentrated sodium hydroxide solution. The eluate contained 95% of the quantity of inhibitor present in the extract in a purity of 60 APU/ml at E280 "* 1,0.
The eluate obtained in the afore-described manner could be freed from higher molecular impurities advantageously by gel chromatography, for example on Sephadex^ G-50 (Deutsche Pharmacia, Frankfurt/M. ) . Thereby, tix@ active substance was obtained with a purity of 140-200 APU/ml at A ¾80 β 1,0 and in a yield of more than 50%, referred to the raw extract.
If desired or required, the proteinase inhibitor so obtained may be subjected to a process for removin pyrogens, adjusted to a therapeutically usual concentration of 250 APU/ml, filtered under sterile conditions and filled into ampuls.
In this form, the proteinase inhibitor may be administered parenterally , in particular intravenously.
EXAMPLES 3 to 11: 5 Liters each of a lung tissue raw extract containing about 6 APU/ml were combined with different amounts of various ion exchangers and worked up essentially as described in Example 2.
The following compilation shows, by way of example, some of the tests carried out. i Ion exchanger Reaction Proteinase conditions inhibitor Example Designation Functional Quantity Ads. Eluant 'yield Purity group g/1 at % APU/ml PH at E280= 1 3 Copolymer Sulfonic 20 8.5 Water/ 92 60 gel acc.to acid 10% NaCl Example 1 pH 3.0 4 Copolymer Sulfonic 20 8.5 Water 81 70 gel acc. to acid pH 12.5 Ex. 1 5 Copolymer Sulfonic 20 8.5 Water/ 82 71 gel acc. to acid 10% NaCl Ex. 1 pH 4.5 6 Copolymer Sulfonic 20 4.5 Water 92 10 gel acc.to acid pH 12.5 Ex. 1 7 Copolymer Sulfonic 20 4.5 Water/ 85 13 gel acc.to acid 10% NaCl Ex. 1 pH 4.5 8 Bio-Rex(R) Sulfonic 2 8.5 Water 90 53 40 *) acid pH 12.5 9 Bio-Rex (R Sulfonic 2 8.5 Water/ 65 52 40 *) acid 20% NaCl pH 7.0 10 AG-MP-50^* Sulfonic 4 8.5 Water 94 32 ac d pH 12.5 11 Bio-Re ^ Phosphonic 6 8.5 Water 75 53 63 *) acid pH 12.5 *) Bio-Rad Laboratories, Richmond, California, U.S.A.
Claims (11)
1. ) A process for the isolation of a polyvalent proteinase inhibitor from animal organs, which comprises adsorbing the inhibitor from its aqueous solutions at pH 0.5 to 10.5 on an ion exchanger containing functional sulfonate or phosphonate groups and eluting it with water or a salt solution at pH 1 to pH 13.
2. ) A process as claimed in claim 1), wherein a copolymer gel of ethene-sulfonate and acryl-amide is used as adsorbant.
3. ) A process as claimed in claims 1 and 2, wherein a gel prepared by copolymerization of ethene-sulfonic acid and acryl-amide in a proportion by weight of 20:1 to 2:1 and subsequent cross-linking with 1 to 15% of formaldehyde is used for the adsorption.
4. ) A process as claimed in claim 1), wherein a polyphenol- Lj~ -sulfonic acid ion exchanger is used as adsorbant.
5. ) A process as claimed in claim 1), wherein a polystyrene/ sulfonic acid ion exchanger, preferably with macro-porous structure, is used as adsorbant.
6. ) A process as claimed in claim 1), wherein a polystyrene/ phosphonic acid ion exchanger is used as adsorbant.
7. ) A process as claimed in claims 1 to 6, wherein the elution is effected at about pH 12.5 with water without addition of salts.
8. ) A process as claimed in claims 1 to 6, wherein the eluting solutions contain increasing amounts of salt with falling pH-value. ^
9. ) A process as claimed in claim 8, wherein, if the copolymer gel of ethene-sulfonate/acryl-amide of claim 3) is used, the elution of the proteinase inhibitor is carried out with a 10% sodium chloride solution at pH 3.0.
10. ) A process as claimed in claim £&) , wherein, if polyphenol- -sulfonic acid ion exchangers, polystyrene-sulfonic acid resins or polystyrene-phosphonic acid resins are used, the elution of the proteinase inhibitor is carried out with a 20% sodium chloride solution at pH 7.0.
11. ) A polyvalent proteinase inhibitor, prepared according to any one of the processes claimed in claims 1 to 10). COHEN ZEDE & SPISBACH P.0. Box 331 1 6 , Tel- A v i v Attorneys for Applicant O ~
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE2406971A DE2406971B2 (en) | 1974-02-14 | 1974-02-14 | Process for obtaining a polyvalent proteinase inhibitor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| IL46587A0 IL46587A0 (en) | 1975-04-25 |
| IL46587A true IL46587A (en) | 1977-06-30 |
Family
ID=5907334
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| IL46587A IL46587A (en) | 1974-02-14 | 1975-02-07 | Process for the isolation of a polyvalent proteinase inhibitor |
Country Status (14)
| Country | Link |
|---|---|
| JP (1) | JPS50111211A (en) |
| AT (1) | AT338414B (en) |
| BE (1) | BE825570A (en) |
| CH (1) | CH610909A5 (en) |
| DE (1) | DE2406971B2 (en) |
| DK (1) | DK145468B (en) |
| FR (1) | FR2261286B1 (en) |
| GB (1) | GB1493960A (en) |
| IE (1) | IE40600B1 (en) |
| IL (1) | IL46587A (en) |
| IT (1) | IT1043948B (en) |
| LU (1) | LU71826A1 (en) |
| NL (1) | NL7501488A (en) |
| SE (1) | SE7501654L (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2527222A1 (en) * | 1982-05-19 | 1983-11-25 | Christine Fougnot | METHOD FOR SEPARATING AND PURIFYING PROTEASES AND ANTIPROTEASES OF BLOOD COAGULATION, AS WELL AS PROTEASE / ANTIPROTEASE COMPLEX |
-
1974
- 1974-02-14 DE DE2406971A patent/DE2406971B2/en not_active Ceased
-
1975
- 1975-02-07 IL IL46587A patent/IL46587A/en unknown
- 1975-02-07 NL NL7501488A patent/NL7501488A/en not_active Application Discontinuation
- 1975-02-12 LU LU71826A patent/LU71826A1/xx unknown
- 1975-02-12 IT IT20216/75A patent/IT1043948B/en active
- 1975-02-13 DK DK52175AA patent/DK145468B/en not_active Application Discontinuation
- 1975-02-13 AT AT107175A patent/AT338414B/en not_active IP Right Cessation
- 1975-02-13 IE IE280/75A patent/IE40600B1/en unknown
- 1975-02-13 JP JP50017517A patent/JPS50111211A/ja active Pending
- 1975-02-13 CH CH174675A patent/CH610909A5/en not_active IP Right Cessation
- 1975-02-14 FR FR7504757A patent/FR2261286B1/fr not_active Expired
- 1975-02-14 GB GB6296/75A patent/GB1493960A/en not_active Expired
- 1975-02-14 SE SE7501654A patent/SE7501654L/ not_active Application Discontinuation
- 1975-02-14 BE BE153395A patent/BE825570A/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| AT338414B (en) | 1977-08-25 |
| FR2261286A1 (en) | 1975-09-12 |
| BE825570A (en) | 1975-08-14 |
| ATA107175A (en) | 1976-12-15 |
| IL46587A0 (en) | 1975-04-25 |
| JPS50111211A (en) | 1975-09-01 |
| CH610909A5 (en) | 1979-05-15 |
| IE40600B1 (en) | 1979-07-04 |
| GB1493960A (en) | 1977-12-07 |
| DK52175A (en) | 1975-10-06 |
| DK145468B (en) | 1982-11-22 |
| SE7501654L (en) | 1975-08-15 |
| FR2261286B1 (en) | 1979-04-06 |
| DE2406971A1 (en) | 1975-08-21 |
| NL7501488A (en) | 1975-08-18 |
| LU71826A1 (en) | 1976-12-31 |
| IT1043948B (en) | 1980-02-29 |
| IE40600L (en) | 1975-08-14 |
| DE2406971B2 (en) | 1979-08-09 |
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