Summary
The production of human gamma interferon as intracellular inclusion bodies in Escherichia coli, which simplified the purification process, is described. An expression plasmid carrying lipoprotein and the tryptophane promoters in tandem was used. Preparation of highly pure interferon was achieved using high resolution chromatography after denaturation and renaturation steps. Structural characteristics of this protein were verified by mass spectrometric analysis. Additional control tests have shown the suitability of the final product for clinical purposes.
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References
Anonymous (1983) Points to consider in the production of new drugs and biologicals by recombinant DNA technology. WHO, Bethesda, Md
Arakawa T, Alton K (1985) Preparation and characterization of recombinant DNA-derivative Hu-Interferon-λ. J Biol Chem 260:14435–14439
Arakawa T, Hsu Y (1985) Structural studies on acid unfolding and refolding of recombinant Hu-interferon gamma. Biochemistry 24:7959–7972
Arakawa T, Hsu Y (1987) Acid unfolding and self-association of recombinant Escherichia coli derived Hu-IFN-λ. Biochemistry 26:5428–5432
Barber M, Bordoli R (1981) Fast atom bombardment of solids as an ion source in mass spectrometry. Nature 293:270–275
Birnboim H, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1524
Box G, Wilson K (1951) On the experimental attainment of optimum conditions. J Roy Stat Soc B 13:1–45
Carlemalm E, Villiger W, Hobot JA, Acetarin JD, Kellenberger (1985) Low temperature embedding with Lowicrylresins: two new formulations and some applications. J Microsc 140:55
Chen A, Anicetti V, Klassen T, Berthold W (1986) A sensitive radioimmunoprecipitation assay for the detection of antibody to recombinant human gamma interferon. Interferon Res 6:313–318
Delgado J, Herrera L, Perez C, Lopez R (1979) Optimization of the selection of auxotrophic mutants in Candida utilis by snail enzyme treatment. Can J Microbiol 25:486–490
Gibson BW, Biemann K (1984) Strategy for the mass spectrometry verification and correction of the primary structures of protein deduced from their DNA sequences. Proc Natl Acad Sci USA 81:1956–1960
Gonzales M, Novoa L, Estrada M, Santos A, Herrera L (1988) An immunoblot technic for insoluble proteins. (eds) Tenth Scientific Seminar, Havana, Cuba
Hanaham D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Knight E (1987) Purification of interferons. Interferon Res 7:501–543
Kung H, Honda S (1986) Purification of recombinant human immune interferon. Methods Enzmyol 119:204–210
Laemmli J (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Langer J, Pestka S (1985) Structure of interferons. Pharmacol Ther 27:371–401
Maniatis T, Fritsch E, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratories, Cold Spring Habor, NY
Martson F (1986) The purification of eukaryotic polypeptides synthesized in Escherichia coli. Biochemistry 240:1
Martson F, LOWE p, Doel P, Schoemaker J, White S, Angal S (1984) Purification of calf prochymosin (prorennin) synthesized in Escherichia coli. Bio/Technology 2:800–804
Morris H, Panico M, Taylor G (1983) Fast atom bombardment of recombinant DNA protein products. Biochem Biophys Res Commun 117:299–305
Nakamura K, Inouye M (1982) Construction of versatile expression coloning vehicles using the lipoprotein gene of Escherichia coli. EMBO J 1:771–777
Nisbet I, Beilharz M, Hertzog P, Tymms M, Linnane A (1985) Single amino acid substitutions at conserved residues of Hu-IFN-λ can affect antiviral specific activity. Biochem Int 11:301–305
Pan Y, Stern A, Familletti P, Khan F (1987) Structural characterization of Hu-IFN-λ. Eur J Biochem 166:145–149
Perez C, Herrera L (1981) Preliminary genetic studies in Candida utilis II. Exp Mycol 5:15–22
Pestka S, Langer J (1987) Interferons and their actions. Ann Rev Biochem 56:727–777
Sedmak J, Grossberg (1981) Approaches to the stabilization of interferons. Texas Rep Biol Med 41:274
Simons G, Renaut E, Allet B, Devos R, Fiers W (1984) High level expression of Hu-IFN-λ in Escherichia coli under control of the pL promoter of bacteriophage lambda. Gene 28:55–64
Steward W, Somer P de, Clercq (1974) Protective effect of anionic detergents on interferons. Biochim Biophys Acta 359:364–373
Steward W, Somer P de, Clercq (1974) Renaturation of inactivated interferons. J Gen Virol 24:567–582
Takao T, Hitouji T, Shimonishi Y, Tanabe T, Inouge S, Inouyi M (1984) Verification of protein sequence by fast atom bombardment-mass spectrometry. Amino-acid sequence of protein S, a development-specific protein of Myxococcus xanthus. J Biol Chem 259:6105–6109
Takao T, Kobayashi M, Nishimura O, Shimonishi Y, (1987) Chemical characterization of recombinant human leukocyte interferon using fast atom bombardment-mass spectrometry. 262:3541–3547
Taylor G, Hoare M, Gray D, Martson F (1986) Bio/Technology 4:553–557
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Perez, L., Vega, J., Chuay, C. et al. Production and characterization of human gamma interferon from Escherichia coli . Appl Microbiol Biotechnol 33, 429–434 (1990). https://doi.org/10.1007/BF00176659
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DOI: https://doi.org/10.1007/BF00176659