EP0165942A1 - PRODUCTION D'INTERFERON $g(a) HUMAIN - Google Patents

PRODUCTION D'INTERFERON $g(a) HUMAIN

Info

Publication number
EP0165942A1
EP0165942A1 EP19850900024 EP85900024A EP0165942A1 EP 0165942 A1 EP0165942 A1 EP 0165942A1 EP 19850900024 EP19850900024 EP 19850900024 EP 85900024 A EP85900024 A EP 85900024A EP 0165942 A1 EP0165942 A1 EP 0165942A1
Authority
EP
European Patent Office
Prior art keywords
ifn
nucleotide sequence
human interferon
polypeptide
sequence
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.)
Withdrawn
Application number
EP19850900024
Other languages
German (de)
English (en)
Inventor
Anthony William Linnane
Phillip Nagley
Sangkot Marzuki
Manfred Werner Beilharz
Ian Thomas Nisbet
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.)
CSL Ltd
Monash University
Original Assignee
Commonwealth Serum Laboratories
Monash University
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 Commonwealth Serum Laboratories, Monash University filed Critical Commonwealth Serum Laboratories
Publication of EP0165942A1 publication Critical patent/EP0165942A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • 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]
    • C07K14/56—IFN-alpha

Definitions

  • This invention relates to a human . interferon- ⁇ and to the production thereof.
  • the invention relates to a complete nucleotide sequence of a human interferon- ⁇ gene, and recombinant DNA molecules comprising this nucleotide sequence, as well as processes utilising said recombinant DNA molecules for producing the human interferon- ⁇ .
  • the human interferons are a group of proteins possessing potent antiviral, antiproliferative and immune response modulating activities (1) .
  • the potential therapeutic value of the interferons together with their limited availability from natural sources, considerable effort has been directed towards the cloning and expression of interferon genes.
  • Three distinct types of interferon, ⁇ , ⁇ and ⁇ , have been described, based on differences in antigenicity and biological characteristics of the molecules (for review, 2) .
  • nucleotide sequence of this aspect of the invention may be obtained from natural, synthetic or semi-synthetic sources; furthermore, the nucleotide sequence may be a naturally-occurring sequence, or may be related by mutation, including single or multiple base substitutions, deletions insertions and inversions, to such a naturally-occurring sequence, provided always that the DNA molecule comprising such a sequence is capable of being expressed as the desired amino acid sequence.
  • the nucleotide sequence may have expression control sequences positioned adjacent to it, such control sequences being derived either from homologous or heterologous sources.
  • the nucleotide sequence of IFN- ⁇ Ml according to this invention is further characterised in having a restriction map substantially as shown in Figure lb.
  • IFN- ⁇ 4a 153 of 189 amino acids
  • ⁇ 4b a complete amino acid sequence for another IFN- ⁇ denoted ⁇ 4b.
  • the amino acid sequences were derived from unpublished nucleotide sequences of clones isolated from the same human gene library as that used in the present report (12) .
  • IFN- ⁇ 4a and IFN- ⁇ 4b are considered to be allelic as they have similar flanking DNA sequences and, on currently published data, have only two amino acid differences (3) .
  • an Alul fragment containing the coding region of the IFN- ⁇ Ml gene has been inserted into the Hindi site of the phage Ml3 mp 11, resulting in a fusion of the IFN- ⁇ Ml gene and the ⁇ -galactosidase gene.
  • E. coli infected with the recombinant M13 phage carrying the fused gene has been cultured and extracts have shown antiviral activity in cytopathic effect inhibition assays. This antiviral activity was completely neutralised by IFN- ⁇ antibodies.
  • Figure la shows the restriction map of the ⁇ Ml PstI fragment containing the IFN- ⁇ Ml gene. Restriction sites are indicated by the symbols ⁇ Q, Pstl ? - ⁇ , EcoRI; f r Hindlll. The hatched area indicates the IFN- ⁇ Ml coding region. The direction of transcription is from left to right.
  • Figure lb shows the restriction map of the ⁇ Ml Rsal fragment containing the IFN- ⁇ Ml gene and the strategy for sequencing the IFN- ⁇ Ml gene. Restriction sites are indicated by the symbols: ⁇ >, Rsal; ⁇ *, Bspl; ⁇ r, Alul; , Sau3AI; ⁇ , EcoRII; ⁇ , Hindlll. Arrowed segments below the map indicate the extent and direction of nucleotide sequence data obtained from M13 subclones. The asterisks indicate the Bspl and EcoRII sites which are present in IFN- ⁇ Ml but absent from both IFN- ⁇ 4a and IFN- ⁇ 4b (see below) .
  • Figure 2 shows the nucleotide and predicted amino acid sequence of IFN- ⁇ Ml.
  • the initiation codon for pre-interferon, the codon for the N-terminal amino acid of the mature interferon and the termination codon are underlined.
  • the putative 'TATA' box is underlined twice-.
  • Figure 3 shows the nucleotide sequence of the M13 recombinant phage M13- ⁇ Ml-Bl in the region of the fusion between the ⁇ -galactosidase gene of Ml3mpll and the IFN- ⁇ Ml gene.
  • Hybridization was carried out using the synthetic oligonucleotide probes (Table 1) . Restriction maps were constructed using standard methods, including-
  • a PstI fragment of the- ⁇ Ml DNA to which the oligonucleotide probes hybridized was subcloned into p ⁇ C9 by standard methods. Following amplification in E. coli the purified PstI fragment was digested with selected restriction enzymes and the resulting fragments were separated by electrophoresis on an agarose gel. The fragments were transferred to nitrocellulose paper, hybridized to specific oligonucleotide probes and a restriction map of the PstI fragment was derived (Fig.la) .
  • the gene was expressed in E. coli.
  • An Alul-fragment of 669 bp (Fig.lb) was cloned into the Hindi site of Ml3mpll and clones with the correct orientation of the Alul fragment were selected by hybridization with oligonucleotides 4 and 5 (Table 1) .
  • Ml3- ⁇ Ml-Bl contained the ⁇ -galactosidase promoter and the N-terminal 15 nu ⁇ leotides of the ⁇ -galactosidase gene coding region, 20 nucleotides of the M13mpll polylinker sequence, and 25 nu ⁇ leotides of the IFN- ⁇ Ml leader sequence followed by the mature IFN- ⁇ Ml coding region (Fig.3) .
  • the 23 amino acid interferon leader would be replaced by a 19 amino acid leader (11 residues of which are non-interferon) , assuming the N-terminal methionine is removed from the ⁇ -galactosidase N-terminus.
  • the interferon activity in both the culture supernatant and in the cell pellet extracts' was completely neutralized by both a polyclonal antiserum against human IFN- ⁇ (Cantell) and a monoclonal anti-human IFN- ⁇ antibody. It may be noted that the level of interferon expression obtained was lower than that previously reported with a M13 vector (20) .
  • Factors which may account for this difference include the intrinsic specific activities of the particular interferons, the specific activity of the product of the particular fused gene constructed here

Abstract

Une molécule d'ADN qui, lors de sa sécrétion, code pour un interféron alpha humain appelé IFN-alphaM1, comporte une séquence de nucléotides essentiellement semblable à celle montrée dans la figure 2. Sont également décrits une molécule d'ADN recombinant, un vecteur ou véhicule de clonage et une cellule hôte, contenant chacun cette séquence de nucléotide. Est également décrit un polypeptide possédant une activité d'interféron alpha humain, comportant une séquence d'acides aminés essentiellement semblable à celle montrée dans la figure 2.
EP19850900024 1983-12-23 1984-12-20 PRODUCTION D'INTERFERON $g(a) HUMAIN Withdrawn EP0165942A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AU29/82 1983-12-23
AU298283 1983-12-23

Publications (1)

Publication Number Publication Date
EP0165942A1 true EP0165942A1 (fr) 1986-01-02

Family

ID=3693481

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19850900024 Withdrawn EP0165942A1 (fr) 1983-12-23 1984-12-20 PRODUCTION D'INTERFERON $g(a) HUMAIN

Country Status (2)

Country Link
EP (1) EP0165942A1 (fr)
WO (1) WO1985002862A1 (fr)

Families Citing this family (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0154186B1 (fr) * 1984-02-10 1992-07-22 Cytoclonal Pharmaceutics Inc. Séparation d'un gène d'interféron et expression de celui-ci
IS1355B6 (is) * 1984-11-12 1989-04-19 Lister Institute Of Preventive Medicine Fjölkjarna kannar
CA2019803C (fr) * 1989-06-29 2000-04-25 Akira Yanai Interferon felin et procede de production
JP4383530B2 (ja) 1996-04-05 2009-12-16 ノバルティス バクシンズ アンド ダイアグノスティックス， インコーポレーテッド 細胞巨大分子合成の阻害が減少したアルファウイルスベクター
EP0935659A1 (fr) 1996-09-17 1999-08-18 Chiron Corporation Compositions et procedes de traitement de maladies intracellulaires
JP2002533124A (ja) 1998-12-31 2002-10-08 カイロン コーポレイション Ｈｉｖポリペプチドの改善された発現およびウイルス様粒子の生成
US7935805B1 (en) 1998-12-31 2011-05-03 Novartis Vaccines & Diagnostics, Inc Polynucleotides encoding antigenic HIV Type C polypeptides, polypeptides and uses thereof
WO2003004620A2 (fr) 2001-07-05 2003-01-16 Chiron, Corporation Polynucleotides codant des polypeptides de type c du vih antigeniques, polypeptides et leurs utilisations
CA2452119C (fr) 2001-07-05 2013-10-15 Chiron Corporation Polynucleotides codant pour des polypeptides antigeniques du vih de sous-type b et/ou de sous-type c, polypeptides et leurs utilisations
AU2003228751A1 (en) 2002-05-01 2003-11-17 Cell Genesys, Inc. Lentiviral vector particles resistant to complement inactivation
WO2006050394A2 (fr) 2004-11-01 2006-05-11 Novartis Vaccines And Diagnostics Inc. Approches combinatoires destinees a produire des reponses immunitaires
US20100196336A1 (en) 2006-05-23 2010-08-05 Dongsu Park Modified dendritic cells having enhanced survival and immunogenicity and related compositions and methods
EP2870238A4 (fr) 2012-07-05 2016-03-09 Ohio State Innovation Foundation Compositions et procédés associés à des vaccins viraux
US11324816B2 (en) 2015-08-31 2022-05-10 Technovax, Inc. Human respiratory syncytial virus (HRSV) virus-like particles (VLPS) based vaccine
WO2017180770A1 (fr) 2016-04-13 2017-10-19 Synthetic Genomics, Inc. Systèmes de réplicon d'artérivirus recombinant et utilisations correspondantes
EP3526332A1 (fr) 2016-10-17 2019-08-21 Synthetic Genomics, Inc. Systèmes réplicons de virus recombinants et leurs utilisations
BR112019011661A2 (pt) 2016-12-05 2020-01-07 Synthetic Genomics, Inc. Composições e métodos para aumentar expressão de gene
EA202091516A1 (ru) 2017-12-19 2020-11-03 Янссен Сайенсиз Айрлэнд Анлимитед Компани Способы и композиции для индукции иммунного ответа против вируса гепатита b (hbv)
US11389531B2 (en) 2017-12-19 2022-07-19 Janssen Sciences Ireland Unlimited Company Methods and apparatus for the delivery of hepatitis B virus (HBV) vaccines
EA202091513A1 (ru) 2017-12-19 2020-09-09 Янссен Сайенсиз Айрлэнд Анлимитед Компани Вакцины против вируса гепатита b (hbv) и их применение
AU2019210189A1 (en) 2018-01-19 2020-08-06 Janssen Pharmaceuticals, Inc. Induce and enhance immune responses using recombinant replicon systems

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CH651308A5 (de) * 1980-07-01 1985-09-13 Hoffmann La Roche Interferone und deren herstellung.
US5098703A (en) * 1982-01-15 1992-03-24 Cetus Corporation Interferon-alpha 76
EP0098864A1 (fr) * 1982-01-15 1984-01-25 Cetus Corporation Interferon alpha 74
CA1217440A (fr) * 1982-08-18 1987-02-03 Michael A. Innis INTERFERON .alpha. 6L
FR2541579B1 (fr) * 1983-02-24 1987-12-18 Inst Organicheskogo Sinteza Ak Interferon n leucocytaire humain et procede d'obtention de celui-ci dans les cellules bacteriennes

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO8502862A1 *

Also Published As

Publication number Publication date
WO1985002862A1 (fr) 1985-07-04

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Inventor name: LINNANE, ANTHONY, WILLIAM

Inventor name: NAGLEY, PHILLIP

Inventor name: BEILHARZ, MANFRED, WERNER

Inventor name: MARZUKI, SANGKOT

Inventor name: NISBET, IAN, THOMAS