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WO1985005619A1 - Nouveau polypeptide et son procede de preparation - Google Patents

Nouveau polypeptide et son procede de preparation Download PDF

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Publication number
WO1985005619A1
WO1985005619A1 PCT/JP1984/000434 JP8400434W WO8505619A1 WO 1985005619 A1 WO1985005619 A1 WO 1985005619A1 JP 8400434 W JP8400434 W JP 8400434W WO 8505619 A1 WO8505619 A1 WO 8505619A1
Authority
WO
WIPO (PCT)
Prior art keywords
lys
ser
arg
leu
phe
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.)
Ceased
Application number
PCT/JP1984/000434
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English (en)
Japanese (ja)
Inventor
Masakazu Kikuchi
Tsutomu Kurokawa
Susumu Honda
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Takeda Pharmaceutical Co Ltd
Original Assignee
Takeda Chemical Industries Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Takeda Chemical Industries Ltd filed Critical Takeda Chemical Industries Ltd
Priority to IL75302A priority Critical patent/IL75302A0/xx
Priority to AU43219/85A priority patent/AU4321985A/en
Priority to PH32351A priority patent/PH22613A/en
Priority to DK248285A priority patent/DK248285A/da
Priority to EP85106885A priority patent/EP0166993A3/fr
Publication of WO1985005619A1 publication Critical patent/WO1985005619A1/fr
Anticipated expiration legal-status Critical
Priority to US07/129,947 priority patent/US4855409A/en
Ceased legal-status Critical Current

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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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/555—Interferons [IFN]
    • C07K14/57—IFN-gamma
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to a novel polypeptide useful as a pharmaceutical and the like, and a method for producing the same.
  • IFN-II r-type interferon
  • the present inventors believe that the above-mentioned disadvantages of the known ri ⁇ ⁇ —r are based on the 2 ⁇ Cys present at the ⁇ terminal, and that the biological activity is equivalent to or higher than that of r IPN-r Thus, the present invention succeeded in producing a new polypeptide which is less likely to cause dimerization and multi-Jt formation, and completed the present invention.
  • the present invention uses the formula
  • X is preferably a bond.
  • Y is preferably Cys-Oln.Qln or Gin, and! It is preferable that ⁇ Gln.
  • Z is Lys or Ly — Arg— Lys— Arg— Ser— Gln-Met-Leu-Phe-Arg-Gly-Arg (H) or Lye-Arg-Lys-Arg— S ⁇ rGIn— Met— Leu -Phe—Arg-01 y—Arg—Arg—A1a-Ser-Gln (H) is preferred.
  • X is a bond
  • ⁇ is Cya-Gln or ⁇ Gln
  • Z is Lye or ⁇ butide (B).
  • X is the bond O
  • Polypeptide (I) has, for example, ATG at the 5 terminus,
  • the region encoding the polypeptide (I ′) may be any region having the nucleotide sequence encoding the above-described polypeptide ( ⁇ ).
  • TGC CAG or CAG as divided I 1 is, those having a 3 ⁇ 4 group sequence represented by AAG CGA AAA AGQ AGT CAG ATO CTQ TTT CGA GGT CGA AGA GCA TCC CAG (V) as Z 1 is preferred.
  • Translation stop codons include T AA, TGA or TAG, with TAA being preferred.
  • the above DNA consists of a signal peptide between the terminal ATG and the DNA encoding polypeptide ( ⁇ ) ⁇ .
  • the above-mentioned DHA preferably has a promoter upstream of ATG, and the promoter may be any promoter that is suitable for the host used for the production of the transformant.
  • the trp promoter in Escherichia coli (eg, 294, W3110, DH1, etc.), the trp promoter, lac promoter, rec A promoter, ⁇ _ promoter, lpp promoter *, etc. Baci- llua subtilis (eg, MI114), SP01 promoter *, SP02 promoter, I »nP promoter, etc.
  • yeast Sacoha-roayces cerevisiae; eg, AH22
  • SV 0 for animal cells eg, cell C 0 S-7, Chinese hamster cell CH 0, etc.
  • animal cells eg, cell C 0 S-7, Chinese hamster cell CH 0, etc.
  • a DNA (brasmid) having an ATG at the 5 'end and a polypeptide ( ⁇ ) downstream thereof and a translation termination codon downstream thereof is a known IFN- produced by chemical synthesis or gene engineering.
  • r-cDNA or IF-rD! TA derived from chromosome can be produced by processing.
  • Brasmid containing (cDNA) pH IT ti 1101! Refer to Reference Example 2 of EPC Publication No. 0103898 (leakage)] as a raw material, and describe a method for producing a D »A-containing brasmid having a region encoding holybebutide ( ⁇ ; Z is a peptide (H)). This is illustrated below.
  • A can be inserted downstream of an appropriate promoter to introduce it into an appropriate host, but if necessary])
  • An S D (Shine and Dalgarno) sequence may be inserted downstream of the promoter.
  • the transformant of the present invention can be prepared using the expression plasmid obtained as described above in a manner known per se [Cohen S. N. et al .; Pro. HatL Acad. Sci. USA,
  • Polypeptide (I) can be produced by culturing the above-described transgenic plant, producing and accumulating polypeptide (I) in the culture, and collecting this.
  • Examples of the medium include ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ 9 medium containing glucose and casamino acid [
  • Culture is usually performed at 15 to 43 for 3 to 24 hours, and if necessary, aeration and / or agitation can be added.
  • cultivation is performed at a low temperature of about 30 to 36, and inactivation of the cits repressor is about 37-: Preferably at 42'C.
  • an agent such as D-mitmycin C or nadixic acid as necessary and to irradiate with ultraviolet rays. Can be.
  • the cells After culturing, the cells are collected by a known method and, for example, gently washed in a loose mouth liquid. After that, the cells are disrupted by, for example, treatment with a protein denaturant (@sonic wave) or treatment with lysozyme or the like, followed by centrifugation. Difficulty * Obtain the supernatant by a known method.
  • Z is a peptide having all 16 amino acids described above.
  • a transformant containing DNA having a region encoding a polypeptide (IT) is cultured, and the action of the protease in the transformant is determined. It can also be produced by purifying under conditions that are easily affected.
  • a protein purification method generally known may be used.
  • an antibody capable of binding to IFN- or polypeptide (I) can be advantageously purified using that antibody column.
  • Example 12 ( ⁇ 2-11 1 1 monoclonal antibody column), Example of filtration 18 (r3-11.1 monoclonal antibody), an antibody column prepared in the same manner as above) Gin-Asp--Pro-Tyr-Vl-Lys-Qlu-Ala-Glu-Asn-Leu-Lye-Lye-yr-Phe-Asn-Ala-Qly-OH Ram [Special Issue 58–2 151 168 No. 1 (January 15th, 1983)] Example of harm to the specification 11 (W! Ir 2-76.53 Monoclonal antibody) Column))) It is.
  • a polypeptide (I) -containing substance is dissolved in an S buffer near a neutral substance, for example, a phosphate buffer solution such as tris phosphate buffer solution, and the antibody activity is reduced. Adsorb to ram. Next, the column is washed with the same buffer solution, and the polypeptide (I) is eluted.
  • the eluate used is a weakly acidic solution, such as vinegar solution, a solution containing polyethylene glycol, and more antibody than polypeptide (I)! A solution containing a peptide that easily binds, a highly concentrated solution, and a combination of these are used.
  • CDMPI Those which do not accelerate the decomposition of butide (I) are preferred.
  • the column eluate should be neutralized with a normal solution]). If necessary]? The purification operation using the above antibody power and antibody can be performed again.
  • the 1-terminal amino acid in the repeptide (I) is (Un), it may be obtained as a mixture with the polypeptide (I) in which ⁇ Gln is present. If necessary, for example, after the above-mentioned purification operation, the polypeptide (I) in which the N-terminal amino acid is Gin can be obtained by heating or treating with diacid (eg, dilute acetic acid).
  • diacid eg, dilute acetic acid
  • polypeptide (I) solution obtained here is subjected to dialysis, if necessary! ? This can be lyophilized into powder. During freeze-drying, stabilizers such as Sovito-mannitol, dextroth, matose and glycerol * can be added.
  • the polypeptide (I) thus obtained has only one or no Cys, it can be obtained as a stable monomer which is less susceptible to dimerization or multimerization than IFN-r in the prior art. In this case, precipitation hardly occurs, and the biological activity of ⁇ -time * decrease is leveraged to be small, so that it can be advantageously used as pharmaceuticals.
  • Helsingborg peptide (I) of the present invention Boripe Buchido having 1 0 7 specific activity of more than in WE scan activity measurement by cytopathic effect inhibition test of vesicular stomatitis Wie scan for ⁇ human amnion-derived WISH cells (VSV) It can be purified to a known level, and it is known that Naturally derived I * 1 * 1 r type 2 IFN) C Salvin et aL, J
  • the polypeptide (I) of the present invention exhibits antiviral, antitumor, cell growth inhibitory and immunopotentiating effects.
  • the polypeptide (I) of the present invention can be mixed with sterilized water, human blood albumin (HSA), physiological saline, and other known physiologically acceptable carriers, and can be parenterally or topically. «Can be given. For example, 100,000 to 1 image cut per adult, preferably 400000 to 400000 cut per day, by intravenous injection or intramuscular injection! ) Can be administered.
  • Formulations containing ⁇ Lipeptide (I) of the present invention may contain other physiologically acceptable active ingredients such as 3 ⁇ 4, diluents, adjuvants, other carriers, buffers, binders, surfactants, and preservatives. May also be contained.
  • Preparations for intraoral administration may be sterile aqueous solutions or suspensions in physiologically acceptable soot, or sterile powders (usually Boribabe) which can be diluted and used in physiologically acceptable diluents. (Obtained by freeze-drying the peptide (I) solution).
  • the preparation containing the ribeptide (I) of the present invention comprises an active ingredient such as IFN-a, IFN- or IFK-r or interleukin 2 such as a lymphokine, which is one part of the polypeptide (I) of the present invention.
  • an active ingredient such as IFN-a, IFN- or IFK-r or interleukin 2 such as a lymphokine, which is one part of the polypeptide (I) of the present invention.
  • the method of releasing the Z-dew as the antiviral activity (IFii-r activity tt) of the polypeptide was carried out in the following section.
  • I-a and crude IFN- r derived from leukocytes were determined using a test to inhibit the cytopathic effect of VSV against FL cell lines derived from human amniotic membrane. determine the value was standard for I ⁇ 1 N one r.
  • Meniwa was mosquito value calculation of Poripe Buchido in ⁇ of interest, always systems of this standard IFN-gamma Sorting by the aforementioned WISH- VSV Atsushi was carried out, and power was calculated from the ratio.
  • Escherichia coli) 294 / pHIT tr 1101 -d 2 was deposited at the Institute for Fermentation, Osaka under the accession number IF 0-143 550 and traded on June 6, 1984. Accession number: PERUP -7658 to the Microbial Industry Research Institute (FR) of the Ministry of Industry
  • Plasmid ptrp 771 (see Reference Example 2 (I) in the above publication) was cleaved with restriction enzymes Clal and Pstl, and the IPN-r gene obtained by cutting the adapter was connected to the downstream of the trp promoter of the DNA fragment obtained.
  • Escherichia coli 294 was transformed with the plasmid pHIT trpl 101-d2 according to the method of Cohen et al.
  • the transformed Escherichia coli (E. coli) 294 / pHITtr101-d2 was obtained.
  • Example 1 (1) in the same manner as IFN one r expression Burasumido PHITtrp 1101 ⁇ AvaH was digested with Pstl, IFN - Avail containing ⁇ gene portion - Pst to preparative 1 k-to DNA fragments min. Oligonucleotide adapter containing a protein synthesis initiation codon synthesized by the above-described triester method with this DNA fragment.
  • the current vector ptr P 7oi [Reference Example 2 of the above-mentioned publication] (im was digested with the restriction enzyme EcoRI, then partially degraded with Clal, and the resulting margin was repaired with DNA volimerase I large fragment. 4
  • the expression vector ptrp781 was constructed using a DNA ligase to form a circular form, the ClaI recognition site closer to the EcoR I recognition site was broken, and the insertion site for the heterologous gene was E (3 ORI sites).
  • the adapter-linked IFN-r gene is inserted downstream of the tributophan promoter of a DNA fragment obtained by digesting Ptrp781 with the restriction enzymes EcoRI and Ps11, and is ligated using T4DNA ligase.
  • I DNA CIV; however I 1 is CAG, Z 1 has a base S column (V)], Boripe Buchido [1 '; however is Gln, Z is expressed Burasumido pHITtrpl encoding Bae Buchido ([pi)] 201—d3 can be constructed ( Figure 2).
  • the plasmid pH ITtrpl 201-d3 is used to transform E. coli 294 according to the method of Cohen et al.
  • Example 1 In the same manner as in Example 1 (I), IFN-r expression plasmid PHIT trpl101 was digested with restriction enzymes Avail and PStl, and Aval-Pstl 1 kb containing the IFN-fit gene portion was digested. Separate the D If A fragment. The D margin generated by the digestion of this DNA fragment with the restriction enzyme Ava! E]]) After filling in using NA polymerase I large fragment, the protein synthesis starting point was chemically synthesized by the Trieste method. Oligonucleotide linkers containing codons
  • the plasmid pHIT trpl 201-d4 is used to transform Escherichia coli 294 according to the method of Cohen et al. (Supra), and the E. coli 294 / pH IT trp 1201 to d47 containing the plasmid is obtained. .
  • the plasmid pt rp78 treated with EcoRI and PstI was ligated.
  • the IFN-r gene was ligated using T4 DNA ligase to construct an IFN-r expression plasmid pHIT trpi 201 (FIG. 4).
  • the oligonucleotide adapter 1 (CATCGATG) containing the protein synthesis initiation codon synthesized by the triester method was used using T4 DNA ligase.
  • the IFN-r gene thus obtained was inserted downstream of the ptrp 77 1 trp promoter cut with Cla I and Pst I obtained in Example 1 ( ⁇ :), and the polypeptide [I; However, the expression plasmid pHITt ⁇ 1201-d4 encoding Y is a bond and Z is a peptide (M)] was constructed (Fig. 5).
  • Escherichia coli 294 was transformed with the plasmid pHIT t ⁇ 1201—d4 according to the method of Cohen et al. (Supra) to obtain a strain containing the plasmid E. coli 294 ZPHIT trp 120 ld4.
  • Example 1 (Surface) Tfi transformant E. coli 294 / pHI trpl 201-d3 was cultured and extracted in the same manner as in Example 2 (I), and the polypeptide [I; A bond or (and) Met, ⁇ is Gin or (and) ⁇ Gln, Z is a peptide (X)]-containing compound.
  • Example 2 (I) The anti-viral activity of the above was determined, and a plant equivalent to that of Example 2 (I) was obtained.
  • Example 1 (II) The transformant obtained in Example 1 (II) was cultured and extracted from E. coli 294 / pHI trpi 201—d4 'in the same manner as in Example 2 (I).
  • the bacterium 5.9- obtained in the same manner as in Example 2 (I) contains 7 M-inoculated guanidine and 2 mil phen-methyl sulfo-fluoride (XI M tris Suspend in loose mouth liquid (pH 7.0) 1, stir at 4 ⁇ 3 for 1 hour * Centrifuge at 10,000 X for 30 minutes to obtain supernatant 2 to obtain io supernatant, 137 sodium sodium oxalate, 2 ⁇ 7 aM Potassium potassium, ai all Lactate (pH 7.4) consisting of lina sodium phosphate and 1.5 all monobasic phosphate (referred to as PBS below) 26 Mor2—11.1, applied at a flow rate of 1 ⁇ / min to the column volume 1.
  • XI M tris Suspend in loose mouth liquid (pH 7.0) 1, stir at 4 ⁇ 3 for 1 hour * Centrifuge at 10,000 X for 30 minutes to obtain supernatant 2 to obtain io supernatant, 137 sodium sodium oxalate, 2
  • this area 2 was previously treated with a 25 mM ammonium phosphate buffer solution (pH 6.0) containing 1 mM ethylenediaminetetraacetic acid, 15 mM sodium chloride, 10 aM cysteine and 2 M guanidine. It is applied to a column of Sepafari S-200 (Famar) made into a flat mouth (2.6 x 9 ⁇ », column capacity 500 W), eluted with the same mouth liquid and has anti-discharge activity. Min 37 W.
  • Example 2 The frozen S-isomer obtained by the method of (I) and ( ⁇ ) was added to 0.1 M each containing 7 M guanidine and 2 mM phenyl methoxide;! Suspend in 3 times the volume of Tris-HCl buffer (pH 7.0), stir at 4 for 1 hour, and centrifuge at 100,000 X for 30 minutes. The upper part is diluted 14 times with PBS and applied to the antibody column ( ⁇ ⁇ 2-11. 1). The column is then washed with (20 BU sodium phosphate gent containing a solution of guanidine dioxate (X 7.0)) and then washed with 20 mM sodium phosphate containing guanidine diacid.
  • Tris-HCl buffer pH 7.0
  • the mixture is shaken with a buffer solution (pH 7.0) to obtain a compound having antiviral activity, and the fraction is prepared in advance using 1 m diethylene diamine tetraacetic acid. 0.15 M sodium chloride, 10 M cysteine
  • the power of Sephatariya S-200 (manufactured by Fal Macia) equilibrated with 25-BM succinic acid-anhydrous acid solution (H 6.0) containing 2M acid guanidine And elute with the same mouth liquid to obtain a tomato having antiviral activity.
  • Example 25 25 frozen cells obtained by the methods of ( ⁇ ), (I) and (Jiang) were each 0.15 0.1 sodium sodium borate (pH 9.5) 1.5 times
  • OMPI Suspend the volume and briefly stir for 1 hour with a 4 wedge, then extract by applying ultrasonic waves 5 times for 30 seconds, and centrifuge at 30,000 ⁇ 9 for 1 hour to obtain a supernatant. The supernatant is mixed with 25 W of the previously washed PBS and slowly stirred at 4 t for 1 hour. Thereafter, the silica gel is packed into a column, washed with 20 to 30 times the column volume of 1 M NaCl, and then with 0.01 M sodium borate containing 0.5 M tetramethylammonium chloride. Elution with mild mouth liquid (pH & 0) yields about 20 fractions showing anti-wisdom activity.
  • Those corresponding to the above 15 Kd are each a peptide [I; X is a bond or (and) Met, Y is Cys ⁇ Gln, Z is Lys], a polypeptide [I; Hand or (and) Met, Y is Gin or (and) Gln, Z is Lys] and polypeptide !: I; X is a bond or (and) Met Y is a bond, Z is Lys] , 17 Kd correspond to the polypeptides [I; where X is a bond or (and) Met, Y is Cy3-Gln, Z is dibutide (H)], polybutide [ I, where X is a bond or (and) Met, Y is Gin or (and) ⁇ Gln, Z is peptide (2)]-and polypeptide [I;
  • WIPO -26- is a bond or (and) Met, Y is a bond, and Z is a peptide (H)].
  • the culture 9.5 Trp -. 8 Mod 3 medium (sterilized (NH 4) 2 HP0 4 5.0 ⁇ / ⁇ , K 2 HP0 4 6.0 ⁇ Z, KH 2 P0 4 4.0 i,
  • the mixture was transferred to a 14-ml glass tank containing Cimapec containing). ⁇ The cells were cultured at 6.6 to 7.0 by adding NH 4 OH. After 13 hours, IAA was added.
  • the growth of the bacterium was maximized 14 hours after the start of the culture, and the antiviral activity at that time was 5 ⁇ 10 6 U.
  • the polypeptide (I) of the present invention has anti-viral, anti-employment and immunological effects, and is stable, so that it can be advantageously used as a pharmaceutical or the like.

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Abstract

Nouveau polypeptide possédant une activité biologique analogue ou supérieure à celle de l'IFN-gamma et subissant difficilement une dimérisation ou une polymérisation, un transformant contenant de l'ADN pouvant coder le nouveau polypeptide, et procédé de préparation du nouveau polypeptide à partir d'un produit de culture du transformant. L'ADN codant le nouveau polypeptide peut être préparé, par exemple, à partir d'un plasmide connu contenant le cADN du gène de l'IFN-gamma, le pHITtrp 1101 ou le pHITtrp 1201. Cet ADN est lié à un vecteur et introduit dans un hôte pour obtenir un transformant. Pour purifier le polypeptide du produit de culture, on utilise une colonne d'anticorps. Le polypeptide peut être utilisé comme agent antiviral ou antinéoplastique.
PCT/JP1984/000434 1984-06-06 1984-09-11 Nouveau polypeptide et son procede de preparation Ceased WO1985005619A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
IL75302A IL75302A0 (en) 1984-06-06 1985-05-24 Novel polypeptide and production thereof
AU43219/85A AU4321985A (en) 1984-06-06 1985-05-31 Gamma interferon analogues
PH32351A PH22613A (en) 1984-06-06 1985-06-03 Novel polypeptide and production thereof
DK248285A DK248285A (da) 1984-06-06 1985-06-03 Polypeptider samt deres fremstilling og anvendelse
EP85106885A EP0166993A3 (fr) 1984-06-06 1985-06-04 Polypeptide et sa production
US07/129,947 US4855409A (en) 1984-06-06 1987-11-23 Novel polypeptides and method of producing same

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
PCT/JP1984/000292 WO1985005618A1 (fr) 1984-06-06 1984-06-06 Procede de preparation de derives de l'interferon
MCPCT/JP84/00292 1984-06-06

Publications (1)

Publication Number Publication Date
WO1985005619A1 true WO1985005619A1 (fr) 1985-12-19

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PCT/JP1984/000434 Ceased WO1985005619A1 (fr) 1984-06-06 1984-09-11 Nouveau polypeptide et son procede de preparation

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Application Number Title Priority Date Filing Date
PCT/JP1984/000292 Ceased WO1985005618A1 (fr) 1984-06-06 1984-06-06 Procede de preparation de derives de l'interferon

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JP (1) JPS615096A (fr)
KR (1) KR860000377A (fr)
WO (2) WO1985005618A1 (fr)
ZA (1) ZA853966B (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5635399A (en) * 1986-04-24 1997-06-03 Chiron Corporation Retroviral vectors expressing cytokines
US5874077A (en) * 1989-10-24 1999-02-23 Chiron Corporation Human til cells expressing recombinant TNF prohormone
US6936695B1 (en) 1982-05-06 2005-08-30 Intermune, Inc. Manufacture and expression of large structural genes

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB8619725D0 (en) * 1986-08-13 1986-09-24 Hoffmann La Roche Homogenous interferon fragments
JP2653061B2 (ja) * 1986-12-27 1997-09-10 武田薬品工業株式会社 新規ポリペプチドおよびその製造法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5928479A (ja) * 1982-07-12 1984-02-15 エフ．ホフマン　―　ラ　ロシュ　アーゲー インタ−フエロン発現ベクタ−
JPS5951792A (ja) * 1982-02-22 1984-03-26 バイオジェン　インコーポレイテッド Ｄｎａ配列、組換ｄｎａ分子およびヒト免疫インタフエロン様ポリペプチドの製造方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU561343B2 (en) * 1981-10-19 1987-05-07 Genentech Inc. Human immune interferon by recombinant dna

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5951792A (ja) * 1982-02-22 1984-03-26 バイオジェン　インコーポレイテッド Ｄｎａ配列、組換ｄｎａ分子およびヒト免疫インタフエロン様ポリペプチドの製造方法
JPS5928479A (ja) * 1982-07-12 1984-02-15 エフ．ホフマン　―　ラ　ロシュ　アーゲー インタ−フエロン発現ベクタ−

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6936695B1 (en) 1982-05-06 2005-08-30 Intermune, Inc. Manufacture and expression of large structural genes
USRE39821E1 (en) 1982-05-06 2007-09-04 Intermune, Inc. Manufacture and expression of large structural genes
US5635399A (en) * 1986-04-24 1997-06-03 Chiron Corporation Retroviral vectors expressing cytokines
US5874077A (en) * 1989-10-24 1999-02-23 Chiron Corporation Human til cells expressing recombinant TNF prohormone

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Publication number Publication date
JPS615096A (ja) 1986-01-10
ZA853966B (en) 1986-01-29
KR860000377A (ko) 1986-01-28
WO1985005618A1 (fr) 1985-12-19

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