FI85979B - FOERFARANDE FOER FRAMSTAELLNING AV ETT REKOMBINANTFRAMSTAELLT INTERFERON UR BAKTERIECELLER. - Google Patents
FOERFARANDE FOER FRAMSTAELLNING AV ETT REKOMBINANTFRAMSTAELLT INTERFERON UR BAKTERIECELLER. Download PDFInfo
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- FI85979B FI85979B FI853324A FI853324A FI85979B FI 85979 B FI85979 B FI 85979B FI 853324 A FI853324 A FI 853324A FI 853324 A FI853324 A FI 853324A FI 85979 B FI85979 B FI 85979B
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- C—CHEMISTRY; METALLURGY
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- C07K14/52—Cytokines; Lymphokines; Interferons
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- C07K14/56—IFN-alpha
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Abstract
Description
1 859791 85979
Menetelmä rekombinanttivalmistetun interferonin eristämiseksi bakteerisoluista - Förfarande för framställning av ett rekombinantframstäl1t interferon ur bakteriecel1er 5 Patenttijulkaisussa EP-A-0.052.861 on kuvattu bakteerisolujen kemiallinen hajotusmenetelmä rekombinanttivalmistetun interferonin, erityisesti IFN-a:n eristämiseksi, jolle menetelmälle on tunnusomaista seuraavat menetelmävaiheet: 10 1. Bakteerisuspensio, erityisesti E.coli-suspensio, jonka bakteerit sisältävät geeniteknologisesti valmistettua IFN, säädetään mineraalihapon avulla pH-arvoon 2,0 - 2,5 baktee-reiden tappamiseksi.A method for isolating recombinantly produced interferon from bacterial cells - A method for the chemical isolation of recombinantly produced interferon, in particular IFN-α. The bacterial suspension, in particular the E. coli suspension containing the genetically engineered IFN, is adjusted to a pH of 2.0 to 2.5 with mineral acid to kill the bacteria.
15 2. Tapetut bakteerisolut erotetaan.15 2. The killed bacterial cells are separated.
3. Saatu hapan biomassa suspendoidaan uudelleen puskuriin, esimerkiksi tris(2-amino-2-hydroksimetyyli-l,3-propaanidi-oli) + natriumkloridi-puskuriin ja säädetään natriumhydrok- 20 sidin avulla pH-arvoon 7,3, jolloin tuhotaan bakteereiden soiuseinämät.3. The resulting acidic biomass is resuspended in a buffer, for example tris (2-amino-2-hydroxymethyl-1,3-propanediol) + sodium chloride buffer, and adjusted to pH 7.3 with sodium hydroxide to destroy the bacterial walls. .
4. Bakteerisolujen erottamisen jälkeen erotetaan liuoksen sisältämä interferoni tunnetuilla menetelmillä ja puhdiste- 25 taan edelleen.4. After separation of the bacterial cells, the interferon contained in the solution is separated by known methods and further purified.
Edelleen on tunnettua, että polypeptidit, kuten interferonit, kestävät vain vähän mekaanisia vaikutuksia (kts. Proceedings of the Society for experimentel Biology and 30 Medicine 146, 249-253 (1974). Tällaisia mekaanisia vaikutuksia esiintyy kuitenkin bakteerisolujen mekaanisessa hajotuksessa tällöin esiintyvien 1eikkausvoimien vuoksi. Tästä syystä ei myöskään tähän mennessä tunneta mitään bakteerisolujen mekaanista hajotusmenetelmää, jonka tarkoi-35 tuksena on eristää geeniteknologisesti muodostettu polypep-tidi, esimerkiksi interferoni, jossa tämä voidaan eristää korkeilla saannoilla.It is further known that polypeptides, such as interferons, have little resistance to mechanical effects (see Proceedings of the Society for Experimental Biology and Medicine 42, 249-253 (1974)), but such mechanical effects occur due to the shear forces then present in the mechanical disruption of bacterial cells. For this reason, no mechanical method for the mechanical lysis of bacterial cells for isolating a genetically engineered polypeptide, for example interferon, is known to date, in which it can be isolated in high yields.
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Esillä olevan keksinnön tavoitteena on siten tuoda esiin rekombinanttivalmistetul1 e interferonille tarkoitettu parempi uuttomenetelmä bakteerisoluista, jossa menetelmässä bakteerisolut hajotetaan mekaanisesti.It is therefore an object of the present invention to provide an improved method of extracting recombinantly produced interferon from bacterial cells, in which method the bacterial cells are mechanically disrupted.
55
Keksinnön mukaisesti tämä tavoite saavutetaan siten, että bakteerisolut, jotka on tapettu mineraalihapoi11 a tunnetuilla menetelmillä (kts. esimerkiksi "Sterilisation, Desinfektion, Konservierung, Chemotherapie - Verfahren, 10 Wirkstoffe, Prufungsverfahren", Dr. K.H. Wallauser'in ja prof. H. Schmidt'in, Georg Thieme Verlag, Stuttgart, 1967, sivut 145 - 161 ja jotka sisältävät rekombinanttivalmistet-tua interferonia, hajotetaan ilman apuaineita suspendoituna vesipitoiseen happoon pH-arvossa 2,0 - 4,0 1eikkausvoimien 15 avulla sen jälkeen erotetaan suspensiossa oleva interferoni tunnetuilla menetelmillä ja puhdistetaan edelleen.According to the invention, this object is achieved by bacterial cells killed by mineral acid methods by known methods (see, for example, "Sterilization, Disinfection, Conservation, Chemotherapy - Verfahren, Wirkstoffe, Prufungsverfahren", Dr. KH Wallauser and Prof. H. Schmidt). Georg Thieme Verlag, Stuttgart, 1967, pp. 145-161 and containing recombinantly produced interferon is decomposed without excipients suspended in aqueous acid at pH 2.0 to 4.0 by shear forces, followed by separation of the interferon in suspension by known methods. and further purified.
Biomassa voidaan suspendoida esimerkiksi mineraalihappoon, kuten laimeaan suolahappoon tai laimeaan vesipitoiseen 20 a 1 kaani happoon, kuten etikkahappoon tai propionihappoon.The biomass can be suspended, for example, in a mineral acid such as dilute hydrochloric acid or in dilute aqueous 20 a 1 kaic acid such as acetic acid or propionic acid.
Bakteereiden soluseinämät hajotetaan mekaanisesti leikkaus-voimien avulla, esimerkiksi homogenisaattorin avulla, parhaiten Ui tra-Turrax-tyyppisen homogenisaattorin avulla.The cell walls of the bacteria are mechanically disrupted by shear forces, for example by means of a homogenizer, preferably by means of a U1 tra-Turrax type homogenizer.
V·: 25 ‘ : Rekombinanttiva1 mi stetun happostabii1 in interferonin, esi merkiksi Q-interferonin, kuten IFN-a2(arg):n eristämiseksi suoritetaan keksinnön mukainen menetelmä erityisen edullisesti pH-arvossa 2,5 - 3,5, parhaiten laimeassa suolahapos-30 sa pH-arvossa 2,5 tai 1-prosenttisessa etikkahapossa.V ·: 25 ': To isolate recombinantly produced acid-stable interferon, for example interferon Q, such as IFN-α2 (arg), the process according to the invention is particularly preferably carried out at pH 2.5 to 3.5, most preferably in dilute hydrochloric acid. 30 sa at pH 2.5 or 1% acetic acid.
Monissa tapauksissa voi edelleen olla edullista, että homo-genisoinnin jälkeen ennen neutralointia lisätään saostusai-netta, esimerkiksi polyetyleeni-imiiniä 0,1 - 0,5 % konsen-35 traatiossa.In many cases, it may still be advantageous to add a precipitant, for example polyethyleneimine, at a concentration of 0.1 to 0.5% after homogenization before neutralization.
3 859793,85979
Homogenisaattorin kierrosluku ja bakteerisolujen hajottamiseen tarvittava aika riippuu käytetyn biomassan tiheydestä. Jos esimerkiksi biomassan tiheys on 1,03 - 1,05, niin homogenisaattorin kierrosluku on tarkoituksenmukaisesti välillä 5 5000 ja 10000 kierr./min. Kun biomassan tiheys on esimer kiksi 1,04, on homogenisaattorin kierrosluku parhaiten välillä 7000 ja 8000 kierr./min. ja hajotusaika 2 minuuttia.The speed of the homogenizer and the time required to lyse the bacterial cells depend on the density of the biomass used. For example, if the density of the biomass is 1.03 to 1.05, then the speed of the homogenizer is suitably between 5,000 and 10,000 rpm. For example, when the biomass density is 1.04, the speed of the homogenizer is best between 7000 and 8000 rpm. and disintegration time 2 minutes.
Keksinnön mukainen menetelmä voidaan parhaiten suorittaa 10 1aboratoriomi tässä I KA-Ultra-Turrax T 45 laitteella ja suuremmassa mitassa Dispax-reaktori11 a 3-6/6, kumpikin Janke & Kunkel KG-yhtiöstä (D-7813 Staufen).The process according to the invention can best be carried out in a laboratory with a KA KA-Ultra-Turrax T 45 and, on a larger scale, a Dispax reactor 11a 3-6 / 6, each from Janke & Kunkel KG (D-7813 Staufen).
Ultra-Turrax T 45 on dispergointi1aite, jossa terä pyörii 15 suurella kierrosnopeudel1 a rakorengasta vasten ja siten rikkoo bakteerisolut suuriturbulenttisessa kentässä. Bakteeri soi \ijen hajottamiseksi pidetään laitetta dekantteri1a-sissa homogenisoitavan mediumin kanssa, jolloin tämä pyörii ympäri pumppuvaikutuksen vuoksi. Kierrosluku on tällöin 20 välillä noin 5000 ja 10000 kierr./min. ja hajotusaika välillä 1 ja 10 minuuttia.The Ultra-Turrax T 45 is a dispersing device in which the blade rotates at a high speed of 15 against the slit ring and thus breaks the bacterial cells in a high turbulent field. To disintegrate the bacteria, the device is kept in a beaker with the medium to be homogenized, which rotates due to the pump effect. The speed is then between about 5,000 and 10,000 rpm. and a disintegration time between 1 and 10 minutes.
Sitä vastoin Dispax-reaktori 3-6/6-homogenisaattori on pakkosyöttöinen kolmivaiheinen homogenisointikammioiden • ’ 25 läpi samalla 1äpivirtausnopeudel1 a. Tällä tavoin saadaan tasaisesti hajonnut bakteerihomogenisaatti . Kierrosluku tässä Dispax-reaktorissa on 3.000 - 8.000 kierr./min. ja syöttöpaine lähtökohdassa enintään noin 2 baria. Tällä homogenisaattori11 a voidaan homogenisoida 50 1 biomassaa 30 1-10 minuutissa.In contrast, the Dispax reactor 3-6 / 6 homogenizer is a forced feed three-stage through the homogenization chambers • '25 at the same flow rate1a. In this way, a uniformly dispersed bacterial homogenate is obtained. The speed of this Dispax reactor is 3,000 to 8,000 rpm. and a supply pressure at the starting point of up to about 2 bar. With this homogenizer11a, 50 l of biomass 30 can be homogenized in 1-10 minutes.
Kuviossa I on esitetty poikkileikkaus Dispax-reaktorin 3-6/6 tapaisesta homogenisaattorista. Tässä kuviossa F on homogenisoitavan täyttöaineksen tulokammio, E on liukuren-35 gastiiviste käyttöaksel issa H; G on tämän käyttöaksel in päälaakeri. Käyttöakseli11 a H on rivissä toisiinsa nähden kolme sylinterin muotoista roottoria C, jotka ovat koske- 4 85979 tuksessa vastaavasti asetettujen staattoreiden D kanssa, jotka on kiinnitetty homogenisaattorin kotelon sisäseinä-miin. Käyttöakselin H päähän on kiinnitetty kuljetusruuvi, joka ulottuu lähtökohdan A sisään. Staattoreiden jäähdyttä-5 miseen käytetään staattoreiden ympäri ulottuvaa jäähdytys-kammiota, jossa on jäähdytysveden syöttöä varten sisääntuloa L ja ulostulo K. Päälaakerin 1iukurengastiivisteestä pitää huolen veden syöttökohdan J kautta jäähdytysvesi, joka on parhaiten korotetussa paineessa, esimerkiksi 1,5 10 barin paineessa. Sylinterin muotoiset roottorit voivat olla tehdyt kaksoisrummuiksi tai kolmoisrummuiksi tai monikerta-rummuiksi, joiden rumpuseinämät ulottuvat staattoreiden vastaavien vastarumpuseinämien välitiloihin. Homogenisoitava neste johdetaan kierukan B ajamana roottoreiden ja 15 staattoreiden välitilojen läpi, jolloin roottoreiden nopea pyöriminen tuhoaa bakteereiden soluseinämät käytettyjen 1 eikkausvoimien vaikutuksesta, ja näin homogenisoitu aines poistuu homogenisaattorista lähtökohdan A kautta.Figure I shows a cross-section of a homogenizer such as a 3-6 / 6 Dispax reactor. In this figure, F is the inlet chamber of the filler to be homogenized, E is the slip-35 seal on the drive shaft H; G is the main bearing of this drive shaft. The drive shaft 11a H is arranged in relation to each other in the form of three cylindrical rotors C which are in contact with stators D, respectively arranged, fixed to the inner walls of the homogenizer housing. Attached to the end of the drive shaft H is a transport screw which extends inside the starting point A. The stator is cooled-5 be utilized to the stators extending around the cooling chamber, which is for supplying the cooling water inlet and outlet, K. L Main bearing 1iukurengastiivisteestä care to keep the water supply of a cooling water which is preferably at elevated pressure, for example 1.5 to 10 bar pressure. Cylindrical rotors can be made as double drums or triple drums or multiple drums, the drum walls of which extend into the intermediate spaces of the corresponding counter-drum walls of the stators. The liquid to be homogenized is passed, driven by a coil B, through the spaces between the rotors and the stators, whereby the rapid rotation of the rotors destroys the bacterial cell walls under the applied shear forces 1, and the homogenized material leaves the homogenizer via the starting point A.
20 Menetelmä voidaan periaatteessa suorittaa myös muilla tunnetuilla mekaanisilla solujen hajotusmenetelmäl1ä, joita on kuvattu esimerkiksi Biochemical Engineering, sivut 3 5 8 f f , l· teoksessa (Second Edition, Academic Press 1973). Saannot .·. : ovat kuitenkin alhaisempia, kun taas keksinnön mukaisessa . 25 menetelmässä käyttämällä Ultra-Turrax T45 tai Dispax-reak- ; tori 3-6/6 homogenisaattoreita saadaan esimerkiksi interfe ronia korkeammalla saannolla käytetystä biomassasta laskettuna kuin mitä tähän mennessä tunnetuilla menetelmillä, mikä oli odottamatonta edellä mainitun tunnetun tekniikan 30 perusteella.The method can in principle also be carried out by other known mechanical cell lysis methods, described, for example, in Biochemical Engineering, pages 3 8 8 f f, 1 (Second Edition, Academic Press 1973). Rules .·. : are, however, lower, while in the invention. 25 using an Ultra-Turrax T45 or Dispax reagent; tori 3-6 / 6 homogenizers are obtained, for example, in a higher yield of interferon from the biomass used than by hitherto known methods, which was unexpected on the basis of the above-mentioned prior art.
Seuraavat esimerkit selventävät keksintöä lähemmin: - - Esimerkki 1 35 E.coli-kloonin HB 101/pFR 33 (kts. EP-A-0.115.613) baktee---· risuspensio, jonka bakteerit sisältävät inhmisinterferoni 5 85979 alfa-2, säädetään mineraalihappojen avulla pH-arvoon 2,0 - 2,5 bakteereiden tappamiseksi. Tapetut bakteerisolut erotetaan suspensiosta ja syväpakastetaan -20°C:ssa.The following examples further illustrate the invention: - Example 1 A bacterial suspension of E. coli clone HB 101 / pFR 33 (see EP-A-0.115.613) containing human interferon 5 85979 alpha-2 is adjusted. with mineral acids to a pH of 2.0 to 2.5 to kill bacteria. The killed bacterial cells are separated from the suspension and deep-frozen at -20 ° C.
5 20 kg erän A syväpakastettua biomassaa lietettiin 250 lit raan 1-prosenttista etikkahappoa 8°C:ssa. Biomassan täydellisen sulamisen jälkeen dispergoitiin kolmivaiheisella homogenisaattori11 a (Dispax-reaktori 3-6/6). Hienojakoiseksi dispergoituun lysaattiin lisättiin saostusapuaineena 10 0,1 % polyety1eeni-imiiniä. Tämän jälkeen pH-arvo muutet tiin 5n natriumhydroksidi11 a arvoon 7,5. Kahden tunnin pituisen uuttoajan jälkeen uutettu biomassa erotettiin 1autasseparaattorin (Westfalia) avulla 1äpivirtausnopeude1 -la 1,5 1/min. Kirkkaasta liuoksesta otettiin näytteet 15 interferoni- ja proteiinipitoisuuden määrittämistä varten ja suoritettiin testit.5 20 kg of deep-frozen biomass from batch A was slurried in 250 liters of 1% acetic acid at 8 ° C. After complete melting of the biomass, a three-stage homogenizer11a (Dispax reactor 3-6 / 6) was dispersed. To the finely dispersed lysate was added 0.1% polyethyleneimine as a precipitation aid. The pH was then adjusted to 7.5 with 5N sodium hydroxide11a. After an extraction time of 2 hours, the extracted biomass was separated by means of a dish separator (Westphalia) at a flow rate of 1.5 l / min. The clear solution was sampled for interferon and protein levels and tested.
Saanto: 46,9 x 106 I.E./g biomassaa.Yield: 46.9 x 106 I.E./g biomass.
Jotta voitaisiin arvioida homogenisaattorin vaikutus inter-20 feronin saantoon, lietetystä biomassasta otettiin 500 ml ennen dispergointia ja uutettiin ilman homogenisointivai-hetta patenttijulkaisun EP-A-0.052.861 mukaisesti. pH-arvon *-· muutos suoritettiin 3n natriumhydroksidi 11 a arvoon 7.3.In order to evaluate the effect of the homogenizer on the yield of inter-20 feron, 500 ml of the slurry biomass was taken before dispersion and extracted without a homogenization step according to EP-A-0.052.861. A change in pH * - · was performed with 7.3 N sodium hydroxide 11a to 7.3.
Tunnin pituisen uuttoajan jälkeen uutetun biomassan lysaat-. 25 ti kirkastettiin 1aboratoriosentrifuugi11 a. Supernatantista otettiin, kuten edellä, näytteet interferoni- ja proteiinipitoisuuksia varten ja suoritettiin testit. Julkaisun EP-A-0.052.861 mukainen saanto: 25,3 x 106 I.E./g biomas-... saa.After an extraction time of one hour, the lysate of the extracted biomass. 25 t were clarified in a laboratory centrifuge11 a. The supernatant was sampled for interferon and protein levels as above and tested. Yield according to EP-A-0.052.861: 25.3 x 106 I.E./g biomass -...
3030
Esimerkki 2Example 2
: : : Valmistettiin esimerkin 1 mukaisesti käyttämällä erän B::: Prepared according to Example 1 using batch B.
. biomassaa.. biomass.
♦ ] 35 Saanto: 21,3 x 10^ I.E./g biomassaa.♦] 35 Yield: 21.3 x 10 ^ I.E. / g biomass.
Julkaisun EP-A-0.052.861 mukainen saanto: 9,7 x 106 I.E./g biomassaa.Yield according to EP-A-0.052.861: 9.7 x 106 I.E. / g biomass.
6 859796 85979
Esimerkki 3Example 3
Valmistettiin esimerkin 1 mukaisesti käyttämällä erän C bi omassaa.Prepared according to Example 1 using batch C bi as its own.
5 Saanto: 13,8 x 10® I.E./g biomassaa.5 Yield: 13.8 x 10® I.E./g biomass.
Julkaisun EP-A-0.052.861 mukainen saanto: 5,1 x 10® I.E./g biomassaa.Yield according to EP-A-0.052.861: 5.1 x 10® I.E./g biomass.
Esimerkki 4 10Example 4 10
Valmistettiin esimerkin 1 mukaisesti käyttämällä 1000 g panoksen BC24 biomassaa, joka oli otettu 1000 ml:aan laimeaa suolahappoa (pH 2,5; 4 ml 37-prosenttista suolahappoa täytettynä vedellä 10.000 ml:ksi) ja dispergoiti in IKA-15 Ultra-Turrax T45 laitteella.Prepared according to Example 1 using a 1000 g batch of BC24 biomass taken up in 1000 ml of dilute hydrochloric acid (pH 2.5; 4 ml of 37% hydrochloric acid filled with water to 10,000 ml) and dispersed in an IKA-15 Ultra-Turrax T45 .
Saanto: 50 x 10® I.E./g biomassaa.Yield: 50 x 10® I.E./g biomass.
Claims (8)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE3432196 | 1984-09-01 | ||
| DE19843432196 DE3432196A1 (en) | 1984-09-01 | 1984-09-01 | NEW MECHANICAL DIGESTIVE METHOD OF BACTERIAL CELLS FOR ISOLATING RECOMBINANTLY PRODUCED PEPTIDES |
Publications (4)
| Publication Number | Publication Date |
|---|---|
| FI853324A0 FI853324A0 (en) | 1985-08-30 |
| FI853324L FI853324L (en) | 1986-03-02 |
| FI85979B true FI85979B (en) | 1992-03-13 |
| FI85979C FI85979C (en) | 1992-06-25 |
Family
ID=6244436
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| FI853324A FI85979C (en) | 1984-09-01 | 1985-08-30 | FOERFARANDE FOER FRAMSTAELLNING AV ETT REKOMBINANTFRAMSTAELLT INTERFERON UR BAKTERIECELLER. |
Country Status (12)
| Country | Link |
|---|---|
| EP (1) | EP0173924B1 (en) |
| JP (1) | JPH078238B2 (en) |
| KR (1) | KR930002886B1 (en) |
| AT (1) | ATE49231T1 (en) |
| DE (2) | DE3432196A1 (en) |
| DK (1) | DK165748C (en) |
| ES (1) | ES8608038A1 (en) |
| FI (1) | FI85979C (en) |
| HU (1) | HU193806B (en) |
| IE (1) | IE58521B1 (en) |
| IL (1) | IL76264A (en) |
| ZA (1) | ZA856640B (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5196323A (en) * | 1985-04-27 | 1993-03-23 | Boehringer Ingelheim International Gmbh | Process for preparing and purifying alpha-interferon |
| US4675387A (en) * | 1985-07-26 | 1987-06-23 | E. I. Du Pont De Nemours And Company | Method for extracting protein with organic acid |
| US4734362A (en) * | 1986-02-03 | 1988-03-29 | Cambridge Bioscience Corporation | Process for purifying recombinant proteins, and products thereof |
| US5831022A (en) * | 1986-02-18 | 1998-11-03 | Hoffmann-La Roche Inc. | Purification of recombinant human IL-1α |
| US4801686A (en) * | 1986-09-04 | 1989-01-31 | Immunex Corporation | Purification of recombinant interleukin-1 |
| US5179199A (en) * | 1986-10-20 | 1993-01-12 | Genzyme Corporation | Protein purification |
| US4912200A (en) * | 1987-05-11 | 1990-03-27 | Schering Corporation | Extraction of granulocyte macrophage colony stimulating factor from bacteria |
| US4958007A (en) * | 1988-05-17 | 1990-09-18 | Schering-Plough Corp. | Extraction of human interleukin-4- from bacteria |
| EP1539798B1 (en) | 2002-09-06 | 2010-11-24 | Genentech, Inc. | Process for protein extraction |
| JP4779537B2 (en) * | 2005-09-28 | 2011-09-28 | パナソニック株式会社 | Vacuum cleaner |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1432039A (en) * | 1972-11-10 | 1976-04-14 | Dai Nippon Sugar Mfg Co Ltd | Method of treating microorganisms |
| JPS5530834A (en) * | 1978-08-25 | 1980-03-04 | Nec Corp | Method of forming ohmic contact in semiconductor pellet |
| US4315852A (en) * | 1980-11-26 | 1982-02-16 | Schering Corporation | Extraction of interferon from bacteria |
| JPS57118798A (en) * | 1980-11-26 | 1982-07-23 | Schering Plough Corp | Obtaining of interferon solution |
| IT1167610B (en) * | 1982-01-19 | 1987-05-13 | Cetus Corp | MULTICLASS HYBRID INTERFRERON, PHARMACEUTICAL COMPOSITION CONTAINING IT AND PRODUCTION PROCESS |
| WO1984002129A1 (en) * | 1982-11-22 | 1984-06-07 | Takeda Chemical Industries Ltd | Human immune interferon protein and process for its preparation |
-
1984
- 1984-09-01 DE DE19843432196 patent/DE3432196A1/en not_active Withdrawn
-
1985
- 1985-08-23 EP EP85110599A patent/EP0173924B1/en not_active Expired - Lifetime
- 1985-08-23 AT AT85110599T patent/ATE49231T1/en not_active IP Right Cessation
- 1985-08-23 DE DE8585110599T patent/DE3575162D1/en not_active Expired - Fee Related
- 1985-08-29 ES ES546528A patent/ES8608038A1/en not_active Expired
- 1985-08-30 DK DK396685A patent/DK165748C/en not_active IP Right Cessation
- 1985-08-30 KR KR1019850006323A patent/KR930002886B1/en not_active IP Right Cessation
- 1985-08-30 IE IE214185A patent/IE58521B1/en not_active IP Right Cessation
- 1985-08-30 HU HU853309A patent/HU193806B/en not_active IP Right Cessation
- 1985-08-30 JP JP60191818A patent/JPH078238B2/en not_active Expired - Lifetime
- 1985-08-30 IL IL76264A patent/IL76264A/en not_active IP Right Cessation
- 1985-08-30 FI FI853324A patent/FI85979C/en not_active IP Right Cessation
- 1985-08-30 ZA ZA856640A patent/ZA856640B/en unknown
Also Published As
| Publication number | Publication date |
|---|---|
| EP0173924A3 (en) | 1988-02-03 |
| JPS6170984A (en) | 1986-04-11 |
| KR930002886B1 (en) | 1993-04-12 |
| HUT39471A (en) | 1986-09-29 |
| JPH078238B2 (en) | 1995-02-01 |
| ES8608038A1 (en) | 1986-06-01 |
| DK165748B (en) | 1993-01-11 |
| IE852141L (en) | 1986-03-01 |
| FI853324A0 (en) | 1985-08-30 |
| IE58521B1 (en) | 1993-10-06 |
| FI85979C (en) | 1992-06-25 |
| IL76264A (en) | 1990-02-09 |
| DK396685D0 (en) | 1985-08-30 |
| DK165748C (en) | 1993-06-14 |
| HU193806B (en) | 1987-12-28 |
| ZA856640B (en) | 1987-05-27 |
| ES546528A0 (en) | 1986-06-01 |
| DE3432196A1 (en) | 1986-03-06 |
| IL76264A0 (en) | 1986-01-31 |
| EP0173924A2 (en) | 1986-03-12 |
| FI853324L (en) | 1986-03-02 |
| DK396685A (en) | 1986-03-02 |
| ATE49231T1 (en) | 1990-01-15 |
| KR860002571A (en) | 1986-04-26 |
| EP0173924B1 (en) | 1990-01-03 |
| DE3575162D1 (en) | 1990-02-08 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MM | Patent lapsed | ||
| MM | Patent lapsed |
Owner name: BOEHRINGER INGELHEIM INTERNATIONAL GMBH |