DE2333834A1 - CARRIER MATERIAL FOR PROTEINS - Google Patents
CARRIER MATERIAL FOR PROTEINSInfo
- Publication number
- DE2333834A1 DE2333834A1 DE19732333834 DE2333834A DE2333834A1 DE 2333834 A1 DE2333834 A1 DE 2333834A1 DE 19732333834 DE19732333834 DE 19732333834 DE 2333834 A DE2333834 A DE 2333834A DE 2333834 A1 DE2333834 A1 DE 2333834A1
- Authority
- DE
- Germany
- Prior art keywords
- water
- proteins
- carrier material
- carrier
- protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 102000004169 proteins and genes Human genes 0.000 title claims description 14
- 108090000623 proteins and genes Proteins 0.000 title claims description 14
- 239000012876 carrier material Substances 0.000 title claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 239000000741 silica gel Substances 0.000 claims description 4
- 229910002027 silica gel Inorganic materials 0.000 claims description 4
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 claims description 3
- 150000004984 aromatic diamines Chemical class 0.000 claims description 3
- 229910010272 inorganic material Inorganic materials 0.000 claims description 3
- 239000011147 inorganic material Substances 0.000 claims description 3
- 239000005373 porous glass Substances 0.000 claims description 3
- 229920003176 water-insoluble polymer Polymers 0.000 claims description 3
- YBRVSVVVWCFQMG-UHFFFAOYSA-N 4,4'-diaminodiphenylmethane Chemical compound C1=CC(N)=CC=C1CC1=CC=C(N)C=C1 YBRVSVVVWCFQMG-UHFFFAOYSA-N 0.000 claims description 2
- 125000003118 aryl group Chemical group 0.000 claims description 2
- HFACYLZERDEVSX-UHFFFAOYSA-N benzidine Chemical compound C1=CC(N)=CC=C1C1=CC=C(N)C=C1 HFACYLZERDEVSX-UHFFFAOYSA-N 0.000 claims description 2
- 239000007795 chemical reaction product Substances 0.000 claims description 2
- 150000004985 diamines Chemical class 0.000 claims description 2
- 239000012460 protein solution Substances 0.000 claims description 2
- 239000007858 starting material Substances 0.000 claims description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical group [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims 2
- WZCQRUWWHSTZEM-UHFFFAOYSA-N 1,3-phenylenediamine Chemical compound NC1=CC=CC(N)=C1 WZCQRUWWHSTZEM-UHFFFAOYSA-N 0.000 claims 1
- KOGDFDWINXIWHI-OWOJBTEDSA-N 4-[(e)-2-(4-aminophenyl)ethenyl]aniline Chemical compound C1=CC(N)=CC=C1\C=C\C1=CC=C(N)C=C1 KOGDFDWINXIWHI-OWOJBTEDSA-N 0.000 claims 1
- 206010012735 Diarrhoea Diseases 0.000 claims 1
- 239000000843 powder Substances 0.000 claims 1
- 235000018102 proteins Nutrition 0.000 description 10
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 239000002245 particle Substances 0.000 description 5
- 239000004365 Protease Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 108090000270 Ficain Proteins 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 108090000631 Trypsin Proteins 0.000 description 3
- 102000004142 Trypsin Human genes 0.000 description 3
- 239000000969 carrier Substances 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000005859 coupling reaction Methods 0.000 description 3
- 229940079919 digestives enzyme preparation Drugs 0.000 description 3
- 235000019836 ficin Nutrition 0.000 description 3
- POTUGHMKJGOKRI-UHFFFAOYSA-N ficin Chemical compound FI=CI=N POTUGHMKJGOKRI-UHFFFAOYSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 108010001078 naringinase Proteins 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- -1 salt. or acetic acid Chemical compound 0.000 description 3
- 239000012588 trypsin Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 description 2
- 239000004952 Polyamide Substances 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- 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 2
- 235000021240 caseins Nutrition 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 108010030923 hesperidinase Proteins 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 description 2
- 229920002647 polyamide Polymers 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000001100 (2S)-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chroman-4-one Substances 0.000 description 1
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 1
- 239000001606 7-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-5-hydroxy-2-(4-hydroxyphenyl)chroman-4-one Substances 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 101710099628 Beta-glucosidase 1 Proteins 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- QUQPHWDTPGMPEX-UHFFFAOYSA-N Hesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(COC4C(C(O)C(O)C(C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-UHFFFAOYSA-N 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- QUQPHWDTPGMPEX-UTWYECKDSA-N aurantiamarin Natural products COc1ccc(cc1O)[C@H]1CC(=O)c2c(O)cc(O[C@@H]3O[C@H](CO[C@@H]4O[C@@H](C)[C@H](O)[C@@H](O)[C@H]4O)[C@@H](O)[C@H](O)[C@H]3O)cc2O1 QUQPHWDTPGMPEX-UTWYECKDSA-N 0.000 description 1
- 239000000440 bentonite Substances 0.000 description 1
- 229910000278 bentonite Inorganic materials 0.000 description 1
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- 102000005936 beta-Galactosidase Human genes 0.000 description 1
- 108010047754 beta-Glucosidase Proteins 0.000 description 1
- 102000006995 beta-Glucosidase Human genes 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- APSNPMVGBGZYAJ-GLOOOPAXSA-N clematine Natural products COc1cc(ccc1O)[C@@H]2CC(=O)c3c(O)cc(O[C@@H]4O[C@H](CO[C@H]5O[C@@H](C)[C@H](O)[C@@H](O)[C@H]5O)[C@@H](O)[C@H](O)[C@H]4O)cc3O2 APSNPMVGBGZYAJ-GLOOOPAXSA-N 0.000 description 1
- IJGRMHOSHXDMSA-UHFFFAOYSA-O diazynium Chemical group [NH+]#N IJGRMHOSHXDMSA-UHFFFAOYSA-O 0.000 description 1
- QGGZBXOADPVUPN-UHFFFAOYSA-N dihydrochalcone Chemical compound C=1C=CC=CC=1C(=O)CCC1=CC=CC=C1 QGGZBXOADPVUPN-UHFFFAOYSA-N 0.000 description 1
- PXLWOFBAEVGBOA-UHFFFAOYSA-N dihydrochalcone Natural products OC1C(O)C(O)C(CO)OC1C1=C(O)C=CC(C(=O)CC(O)C=2C=CC(O)=CC=2)=C1O PXLWOFBAEVGBOA-UHFFFAOYSA-N 0.000 description 1
- 229940110377 dl- arginine Drugs 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- VUYDGVRIQRPHFX-UHFFFAOYSA-N hesperidin Natural products COc1cc(ccc1O)C2CC(=O)c3c(O)cc(OC4OC(COC5OC(O)C(O)C(O)C5O)C(O)C(O)C4O)cc3O2 VUYDGVRIQRPHFX-UHFFFAOYSA-N 0.000 description 1
- QUQPHWDTPGMPEX-QJBIFVCTSA-N hesperidin Chemical compound C1=C(O)C(OC)=CC=C1[C@H]1OC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]4[C@@H]([C@H](O)[C@@H](O)[C@H](C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-QJBIFVCTSA-N 0.000 description 1
- 229940025878 hesperidin Drugs 0.000 description 1
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- DFPMSGMNTNDNHN-ZPHOTFPESA-N naringin Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC=2C=C3O[C@@H](CC(=O)C3=C(O)C=2)C=2C=CC(O)=CC=2)O[C@H](CO)[C@@H](O)[C@@H]1O DFPMSGMNTNDNHN-ZPHOTFPESA-N 0.000 description 1
- 229940052490 naringin Drugs 0.000 description 1
- 229930019673 naringin Natural products 0.000 description 1
- ARGKVCXINMKCAZ-UHFFFAOYSA-N neohesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(CO)O3)OC3C(C(O)C(O)C(C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UHFFFAOYSA-N 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 150000004760 silicates Chemical class 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
- C12N11/096—Polyesters; Polyamides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
- C12N11/12—Cellulose or derivatives thereof
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Inorganic Chemistry (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Peptides Or Proteins (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
Die Erfindung betrifft ein Trägermaterial für Proteine sowie wasserunlösliche Proteinpräparate und ein Verfahren zu deren Herstellung.The invention relates to a carrier material for proteins and water-insoluble protein preparations and a method for their manufacture.
Bs wurde gefunden, dass man Träger zur Bereitung wasserunlöslicher Protein-, insbesondere Enzympräparate,dadurch erhalten kann, dass man ein wasserunlösliches Polymer oder, vorzugsweise, ein wasserunlösliches anorganisches, reaktive Säuerst of funkt ionen enthaltendes Material mit einem aromatischen Diamin behandelt und das Reaktionsprodukt diazotiert. Durch diesen Prozess werden die Trägerpartikel mit einer unablösbaren Ochicht, die aktive Diazoniumgruppen enthält, versehen. Die so erhaltenen Träger liefern bei der Behandlung mit Proteinlösungen wasserunlösliche Proteinpräparate.It has been found that carriers can be used to prepare water-insoluble Protein preparations, in particular enzyme preparations, can be obtained by mixing a water-insoluble polymer or, preferably, a water-insoluble inorganic, reactive acid-containing material with an aromatic Treated diamine and diazotized the reaction product. Through this process, the carrier particles become indelible Ochicht, which contains active diazonium groups, provided. The so The carriers obtained give water-insoluble protein preparations on treatment with protein solutions.
309885/1268309885/1268
Beispiele vcn anorganischen Materialien, die für die Herstellung des erfindungsgemässen Trägers in Betracht kommen sind Silikate und silikathaltige Materialien, z.B. Sand, Steine, Silicagel oder Kieselgur; Bentonit, Vollastonit oder Glas; aber auch Metalloxide wie Aluminiumoxid und Hydroxylapatit.Examples of inorganic materials used for the Preparation of the carrier according to the invention come into consideration are silicates and materials containing silicate, e.g. sand, stones, silica gel or kieselguhr; Bentonite, solid astonite or Glass; but also metal oxides such as aluminum oxide and hydroxyapatite.
Als Beispiele geeigneter Polymerer seien Polyamide genannt. Auch Cellulose, insbesondere Baumwolle, z.B. in Form von Watte ist geeignet. Examples of suitable polymers are polyamides. Cellulose, in particular cotton, e.g. in the form of wadding, is also suitable.
Beispiele von aromatischen Diaminen sind 1,2-Phenylen— diamin, 4,4'-Diamino-diphenylinethant Benzidin, und 4-,4'-DiSnJiHO-stilben. Grundsätzlich ist für die Herstellung der erfindungsgemässen Träger jedes aromatische Diamin geeignet.Examples of aromatic diamines are 1,2-phenylenediamine, 4,4'-diamino-diphenylinethane t benzidine, and 4-, 4'-DiSnJiHO-stilbene. In principle, any aromatic diamine is suitable for the production of the carriers according to the invention.
Zur Herstellung des erfindungsgemässen Trägenaaterials werden die als Ausgangsstoff verwendeten Partikel zunächst mit einer Lösung des Amins, zweckmässig unter leichtem Erwärmen, behandelt. Danach diazotiert man die so erhaltenen Produkte in an sich bekannter Weise, z.B. mit Nitrit und Säure, wie Salz— oder Essigsäure, zweckmässig bei Temperaturen zwischen 0° und Zimmertemperatur.For the production of the carrier material according to the invention the particles used as the starting material are first treated with a solution of the amine, expediently with gentle heating, treated. The products thus obtained are then diazotized in a manner known per se, e.g. with nitrite and acid, such as salt. or acetic acid, expediently at temperatures between 0 ° and room temperature.
Zur Kupplung des Proteins an den Träger behandelt man den letzteren mit einer Lösung des Proteins, wobei man das Verhältnis Protein:Träger zweckmässig auf etwa 1-50 Milligramm pro Gramm bemisst. Die Behandlung wird zweckmässig in einer Pufferlösung, z.B. in Phosphatpuffer, bei pH 5-7,8, vorzugsweise bei pH 7,5 vorgenommen.To couple the protein to the carrier, the latter is treated with a solution of the protein, the ratio of protein: carrier being expediently measured at about 1-50 milligrams per gram. The treatment is expediently carried out in a buffer solution, for example in phosphate buffer, at pH 5-7.8, preferably at pH 7.5.
Als Proteine kommen für die Kupplung an den erfindungsgemässen Träger Antigene, Antikörper, Inhibitoren, wie Tryptininhibitoren und insbesondere Enzyme in Betracht. Beispiele von Enzymen, von denen erfindungsgemäss unlösliche Enzympräparate hergestellt werden können, sind Naringinase, Hesperidinase, ß-Galactosidase, ß-Glucosidase, Trypsin, Bromalin, Ficin, Papain, oc-Amylase, Amyloglucosidaae und Urease.The proteins used for coupling to the carrier according to the invention are antigens, antibodies, inhibitors, such as Tryptin inhibitors and especially enzymes into consideration. Examples of enzymes, of which according to the invention insoluble Enzyme preparations that can be made are naringinase, hesperidinase, ß-galactosidase, ß-glucosidase, trypsin, bromaline, Ficin, papain, oc-amylase, amyloglucosidaae and urease.
309885/1268309885/1268
Die nachstehenden Beispiele erläutern die Erfindung:The following examples illustrate the invention:
1 g Silicagel (zur Chromatographie) wurde zu einer Lösung von 200 mg 4,4'-Diaminodiphenylmethan in 10 ml trockenem Benzol gegeben. Das Gemisch wurde 5 Minuten unter gelegentlichem Schütteln auf dem Wasserbad (80°) erwärmt, sodann abgenutscht, bis alles Benzol entfernt war. Der Rückstand wurde in 20 ml eiskaltes Wasser gegeben, unter Rühren mit 300 mg Natriumnitrit und sodann mit verdünnter Salzsäure (20 Vol-$) versetzt, bis das pH 1-2 betrug. Danach wurde 10 Minuten bei 0° gerührt, das pH durch Zusatz von 1 N Natronlauge auf pH 5 eingestellt und der Ueberstand abdekantiert. Der Rückstand wurde mehrmals mit eiskaltem Wasser, das durch Zusatz von Salzsäure auf pH 4-5 gebracht worden war, gewaschen, bis der Ueberstand beim Dekantieren farblos war. Die so erhaltenen Partikel wurden unmittelbar für die Enzymkupplung verwendet. Tüne· Lagerung in Pliosphatpuffer pH 6,5 bei 4° während 24 Stunden zeigte keinen merklichen Verlust an Kupplungskapazität.1 g of silica gel (for chromatography) was added to a solution of 200 mg of 4,4'-diaminodiphenylmethane in 10 ml of dry benzene. The mixture was heated on the water bath (80 °) for 5 minutes with occasional shaking, then suction filtered until all of the benzene was removed. The residue was poured into 20 ml of ice-cold water, 300 mg of sodium nitrite and then dilute hydrochloric acid (20% by volume) were added with stirring until the pH was 1-2. The mixture was then stirred at 0 ° for 10 minutes, the pH was adjusted to 5 by adding 1N sodium hydroxide solution and the supernatant was decanted off. The residue was washed several times with ice-cold water, which had been brought to pH 4-5 by adding hydrochloric acid, until the supernatant was colorless on decanting. The particles obtained in this way were used directly for the enzyme coupling. T TSOW · Storage in Pliosphatpuffer pH 6.5 at 4 ° for 24 hours showed no significant loss of coupling capacitance.
In der gleichen Weise wurden Trägermaterialien auf der Basis von porösem Glas (Partikelgrösse 40-200 mesh US Standard, Porengrösse 10-200 nir) Aluminiumoxid, Cellulose und Polyamid hergestellt .In the same way, support materials based on porous glass (particle size 40-200 mesh US standard, Pore size 10-200 nir) made of aluminum oxide, cellulose and polyamide .
1 g des gemäss Beispiel 1 hergestellten Trägermaterials wurden zu 25 ml einer eiskalten Enzymlösung (-siehe Tabelle) in 0,05 M Phosphatpuffer pH 6,5-7,8 (Proteingehalt 1-8 mg), gegeben und bei 4° 30 Minuten bis 20 Stunden leicht geschüttelt. Danach wurde filtriert, die Partikel wurden dreimal mit j>e 50 ml 1 H Kaliumchloridlosung gewaschen, in 20 ml 0,05 M Citrat- oder Phosphatpuffer pH 6,5 suspendiert und bei 4° aufbewahrt. Die1 g of the carrier material prepared according to Example 1 were added to 25 ml of an ice-cold enzyme solution (see table) in 0.05 M phosphate buffer pH 6.5-7.8 (protein content 1-8 mg), given and gently shaken at 4 ° for 30 minutes to 20 hours. Thereafter was filtered, the particles were washed three times with 50 ml of 1 H Washed potassium chloride solution in 20 ml of 0.05 M citrate or Phosphate buffer pH 6.5 suspended and stored at 4 °. the
309885/1263309885/1263
Aktivität der so erhaltenen Enzympräparate ist der nachstehenden Tabelle zu entnehmen:Activity of the enzyme preparations thus obtained is as follows See table:
309885/1263309885/1263
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KJ CD 00KJ CD 00
Enzymmenge
[mg Protein] [Einheiten]Deployed
Amount of enzyme
[mg protein] [units]
8
192
384
10,4
1,2
5,5
1,1
8,83.4
8th
192
384
10.4
1.2
5.5
1.1
8.8
pro g Träger
Einheitenactivity
per g of carrier
units
9,4
2,1
0,44
0,35
1,761.8
9.4
2.1
0.44
0.35
1.76
Amyloglucosidase
Hesperidinase
Ficin
Trypsin
Bromalin
ß-GlucosidaseNaringinase
Amyloglucosidase
Hesperidinase
Ficin
Trypsin
Bromaline
β-glucosidase
2,3
4
8
2
2
2,5
2
21
2.3
4th
8th
2
2
2.5
2
2
2,4
50
90
4,6
0,38
1,9
0,32
2,31.9
2.4
50
90
4.6
0.38
1.9
0.32
2.3
Stärke
Hesperidin-dihydrochalcon
Casein
N-Benzoy1-dl-arginin -4-nitro-anilid
Casein
N-Nitrophenyl-glyqopyranosidNaringin
strength
Hesperidin dihydrochalcone
Casein
N-Benzoy1-dl-arginine -4-nitro-anilide
Casein
N-nitrophenyl glyqopyranoside
OO
OO
OO
CO
OOas above
OO
OO
OO
CO
OO
Amyloglucosidase
Trypsin
Ficin
Bromalin
ß-CHucosidaseNaringinase
Amyloglucosidase
Trypsin
Ficin
Bromaline
β-CHucosidase
2
2,5
2
2
23
2
2.5
2
2
2
90
5,5
1,2
1,1
8,810
90
5.5
1.2
1.1
8.8
in ι in ι
Claims (1)
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CH1042272A CH579533A5 (en) | 1972-07-12 | 1972-07-12 |
Publications (1)
Publication Number | Publication Date |
---|---|
DE2333834A1 true DE2333834A1 (en) | 1974-01-31 |
Family
ID=4363063
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DE19732333834 Pending DE2333834A1 (en) | 1972-07-12 | 1973-07-03 | CARRIER MATERIAL FOR PROTEINS |
Country Status (14)
Country | Link |
---|---|
JP (1) | JPS4959697A (en) |
AT (1) | AT326813B (en) |
AU (1) | AU5578973A (en) |
BE (1) | BE802236A (en) |
CH (1) | CH579533A5 (en) |
DE (1) | DE2333834A1 (en) |
FR (1) | FR2192113B1 (en) |
GB (1) | GB1444395A (en) |
HU (2) | HU167605B (en) |
IL (1) | IL42259A (en) |
IT (1) | IT990560B (en) |
NL (1) | NL7309663A (en) |
SE (1) | SE7309771L (en) |
ZA (1) | ZA733252B (en) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CH579098A5 (en) * | 1973-02-22 | 1976-08-31 | Givaudan & Cie Sa | |
FR2417514A1 (en) * | 1978-02-16 | 1979-09-14 | Rhone Poulenc Ind | Enzymes immobilised on inorganic supports - by attaching to bridging molecule bonded to support by an ester linkage |
DE2963866D1 (en) * | 1978-02-16 | 1982-11-25 | Rhone Poulenc Spec Chim | Support-enzyme complexes and method for preparing them |
DE3311889A1 (en) * | 1983-03-31 | 1984-10-11 | Byk-Mallinckrodt Chemische Produkte Gmbh, 6057 Dietzenbach | METHOD FOR IRREVERSIBLE BINDING OF PROTEIN TO POLYSTYROL SURFACES WITH THE PRESERVATION OF BIOLOGICAL ACTIVITY, POLYSTYROL SURFACES OBTAINED AND THEIR USE |
IL83451A (en) * | 1987-08-06 | 1991-06-10 | Univ Ramot | Stabilized water soluble enzymes and a method for their preparation |
-
1972
- 1972-07-12 CH CH1042272A patent/CH579533A5/xx not_active IP Right Cessation
-
1973
- 1973-05-11 IT IT24005/73A patent/IT990560B/en active
- 1973-05-14 ZA ZA733252A patent/ZA733252B/en unknown
- 1973-05-14 IL IL42259A patent/IL42259A/en unknown
- 1973-05-14 AT AT420673A patent/AT326813B/en not_active IP Right Cessation
- 1973-05-16 AU AU55789/73A patent/AU5578973A/en not_active Expired
- 1973-05-18 HU HUGI191A patent/HU167605B/hu unknown
- 1973-05-18 HU HUGI216A patent/HU168032B/hu unknown
- 1973-07-03 DE DE19732333834 patent/DE2333834A1/en active Pending
- 1973-07-11 FR FR7325371A patent/FR2192113B1/fr not_active Expired
- 1973-07-11 JP JP48078241A patent/JPS4959697A/ja active Pending
- 1973-07-11 SE SE7309771A patent/SE7309771L/sv unknown
- 1973-07-11 NL NL7309663A patent/NL7309663A/xx unknown
- 1973-07-12 BE BE133381A patent/BE802236A/en unknown
- 1973-07-12 GB GB3328873A patent/GB1444395A/en not_active Expired
Also Published As
Publication number | Publication date |
---|---|
IL42259A0 (en) | 1973-07-30 |
BE802236A (en) | 1974-01-14 |
ZA733252B (en) | 1974-03-27 |
ATA420673A (en) | 1975-03-15 |
JPS4959697A (en) | 1974-06-10 |
HU167605B (en) | 1975-11-28 |
NL7309663A (en) | 1974-01-15 |
AU5578973A (en) | 1974-11-21 |
AT326813B (en) | 1975-12-29 |
IT990560B (en) | 1975-07-10 |
FR2192113A1 (en) | 1974-02-08 |
HU168032B (en) | 1976-02-28 |
CH579533A5 (en) | 1976-09-15 |
IL42259A (en) | 1976-09-30 |
FR2192113B1 (en) | 1976-11-12 |
GB1444395A (en) | 1976-07-28 |
SE7309771L (en) | 1974-01-14 |
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