Production of interferon

• US 5,468,608 A
• Filed: 09/28/1982
• Issued: 11/21/1995
• Est. Priority Date: 11/21/1979
• Status: Expired due to Term

• Current Assignee: Yeda Research and Development Co. Ltd. (Weizmann Institute of Science)
• Original Assignee: Yeda Research and Development Co. Ltd. (Weizmann Institute of Science)
• Inventors: Revel, Michel, Tiollais, Pierre
• Primary Examiner(s): Martinell, James
• Application Number:
  US06/425,935
• Time in Patent Office: 4,802 Days
• Field of Search: 435/68, 435/70, 435/91, 435/172, 435/253, 435/317, 435/69.51, 435/91.1, 435/172.3, 435/320.1, 435/252.3-252.35, 435/811, 536/27, 536/23.52, 935/13, 935/21, 530/351
• US Class Current: 435/91.1
• CPC Class Codes: C07K 14/5412 IL-6
• CPC Class Codes: C07K 14/565 IFN-beta

Abstract

The present invention relates to a process to isolate genetic material (DNA) containing the nucleotide sequence coding for interferon in human fibroblastic cells which comprises cultivating cells producing interferon when exposed to an inducer of interferon, exposing same to such inducer, extracting messenger RNA from said induced cells, purifying the interferon messenger RNA, transcribing the messenger RNA into DNA and cloning the DNA in a suitable vector. Preferred cells are human diploid foreskin cells. The invention further relates to a process for engineering a bacterial strain to produce interferon polypeptide which comprises introducing a cloned interferon DNA into a suitable vector-carrier. A preferred vector-carrier is E. coli. The invention also relates to the mRNA of human interferon in highly purified form, to the mRNA of human interferon in β1 highly purified form, to the mRNA of human interferon in β2 highly purified form, to the DNA coding for a polypeptide having interferon activity, insertable in a vector, such as plasmid pBR322, and also to human interferon β1 in highly purified form, and human interferon β2 in highly purified form.

Claims

1. An mRNA which consists essentially of an mRNA which is translatable to human fibroblast interferon β

2 polypeptide.

2. A process for obtaining an mRNA population which includes mRNAs containing the nucleotide sequence translatable to human interferon-β

2, and substantially excluding mRNAs containing the nucleotide sequence translatable to human interferon-β

1, comprising;

a) cultivating human fibroblast cells capable of producing fibroblast interferon-β

when exposed to an inducer of interferon;

b) exposing said host cells to a said inducer of interferon;

c) extracting messenger RNAs from said induced host cells; and

d) purifying said messenger RNAs to obtain the fraction thereof having about 1,250-1,350 nucleotides and corresponding to RNA sedimenting at 14S in sucrose gradient sedimentation.

3. A process in accordance with claim 2, wherein said purifying step comprises subjecting said mRNAs to sucrose gradient sedimentation and recovering the RNAs sedimenting at 14S.

4. A process for obtaining an mRNA population which includes mRNAs containing the nucleotide sequence translatable to human interferon-β

1, and substantially excluding mRNAs containing the nucleotide sequence translatable to human interferon-β

2, comprising;

a) cultivating human fibroblast host cells capable of producing fibroblast interferon-β

when exposed to an inducer of interferon;

b) exposing said host cells to a said inducer of interferon;

c) extracting messenger RNAs from said induced host cells; and

d) purifying said messenger RNAs to obtain the fraction thereof having about 900-1,000 nucleotides and corresponding to RNA sedimenting at 11S in sucrose gradient sedimentation.

5. A process in accordance with claim 4, wherein said purifying step comprises subjecting said mRNAs to sucrose gradient sedimentation and recovering the RNA sedimenting at 11S.