Large-Scale, One-Step Purification of Murine Interferon-Beta Using a Monoclonal Antibody

Vonk, W. P.; Trapman, Jan

doi:10.1089/jir.1983.3.169

Cited by 8 publications

(9 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The specific activity of the fraction eluted from the antibody column (3.3 x 10' units/mg protein) was lower than that reported by Vonk and Trapman (4.9 x 10' U/mg protein) [32]. This might be due to contaminating proteins detected on polyacrylamide gels after silver staining (Fig.…”

Section: Mu-ifn-fi Purijkationmentioning

confidence: 54%

See 1 more Smart Citation

Purification and carbohydrate structure of natural murine interferon‐β

Civas

Fournet

Coulombel

et al. 1988

European Journal of Biochemistry

View full text Add to dashboard Cite

Mouse interferon$ (Mu-IFN-P) induced in C-243 cells with Newcastle disease virus was purified in four steps including ammonium sulfate fractionation, DEAE-cellulose, monoclonal Mu-IFN-P antibody affinity and Mono-S cation-exchange chromatographies. Specific activity of the purified Mu-IFN-P ranged over 1.1 -1.4 x lo9 NIH units/mg protein. This preparation was submitted to pronase digestion and gel filtration on Fractogel TSK HW-40. The permethylated and acetylated glycopeptide fraction was analyzed by chemical-ionization (ammonia) mass spectrometry. The major glycopeptide is composed of Gal, Man, GlcNAc and NeuAc with a molar ratio of 2.0: 3.6: 3.4: 0.5. The GLC pattern of methyl derivatives obtained by methanolysis and acetylation of fully methylated glycopeptide identified 2,3,4,6-tetra-O-methylgalactose; 3,4,6-tri-O-methyl-mannose; 2,3,4-and 2,4,6-tri-0-methyl galactose; 2,4,di-O-methyl mannose and 3,6-di-O-rnethylglucosamine.These results when compared with data on N-glycans suggest the following structure for the carbohydrate moiety of Mu-IFN-P:Murine interferons a and fl are released from various cells in response to virus infection [l-41 or upon induction by double-stranded polyribonucleotide [5]. Murine interferon-? is produced by lymphocytes exposed to a variety of antigens or mitogen stimulations [6]. These three types of IFN are glycosylated proteins which are distinct in their antigenicityThe potential therapeutic application of IFNs for treating viral infections or tumor proliferation [lo] established a priority in the cloning of human IFNs [ll]. Nevertheless, interferon's antiproliferative and pharmacodynamic effects requiring in vivo experimentation had to be performed in well defined systems such as isogenic mouse strains.For these reasons, recombinant murine IFNs have been obtained and their primary structure have been determined (see [12] for review). These studies showed that Mu-IFN-a polypeptides are the products of at least ten different genes which seem to be highly conserved during evolution [13]. Sequence analysis of seven of them indicated that the mature Mu-IFN-a proteins vary in size from 162 to 167 amino acids; all except two possess a single N-glycosylation site (Asn-AlaThr) at position 78.[7-91.Correspondence to J. Doly, Biologie Moleculaire des Interfkrons, Institut de Recherches sur le Cancer, 7, Rue Guy-Mocquet, F-94802 Villejuif Cedex, FranceAbbreviations. Mu-IFN, murine interferon; Hu-IFN, human interferon; MEM, minimal essential medium; NDV, Newcastle disease virus; VSV, vesicular stomatitis virus; PBS, 0.02 M phosphate buffer pH 7.3 and 0.15 NaCl; CHO, Chinese hamster ovary. M a n LY I

show abstract

Section: Mu-ifn-fi Purijkationmentioning

confidence: 54%

“…Fractions of 2 ml were collected when the bound material was eluted with 0.1 M citrate buffer pH 2.2 containing 1 M NaCl as described by Vonk and Trapman [32] and assayed for antiviral activity. The column was regenerated with PBS pH 8,0.02% NaN3.…”

Section: Antibody Affinity Chromatographymentioning

confidence: 99%

Purification and carbohydrate structure of natural murine interferon‐β

Civas

Fournet

Coulombel

et al. 1988

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…Conventional MulFN-c~ (subspecies) preparations were prepared as described previously (Vonk & Trapman, 1983;Lemson et al, 1984). Conventional hamster interferon was obtained from the supernatant of CHO cells induced with Sendal virus.…”

Section: Methodsmentioning

confidence: 99%

Constitutive Expression of a Murine Interferon Alpha Gene in Hamster Cells and Characterization of Its Protein Product

et al. 1985

Journal of General Virology

Self Cite

View full text Add to dashboard Cite

SUMMARYThe coding part of a murine interferon alpha (MuIFN-c0 gene was cloned into an expression plasmid containing the simian virus 40 early promoter and the rabbit ~-globin polyadenylation signal. This construct was transfected into Chinese hamster ovary cells, together with a plasmid containing the Ecogpt gene as a selection marker. Resulting colonies were assayed for constitutive interferon production and analysed for integration of MulFN-ct genes. There was no obvious correlation between the number of genes integrated and the amount of interferon produced. The highest producer, designated CHO-pSVIOEF-3, contained four copies of the mouse gene and constitutively secreted up to 100000 International Units of interferon per ml per day. The MulFN-ct subspecies produced by this clone was characterized by analysis of its antiviral activity on heterologous cells, heparin-Sepharose affinity chromatography and chromatofocusing. The results obtained indicate that it is identical or closely related to a minor component present in conventional MulFN-~ preparations.

show abstract

“…We quantitatively separated MulFN-~ from MulFN-fl by affinity chromatography over a monoclonal (EB3) anti-MulFN-fl antibody column. This monoclonal antibody was previously shown to be able to bind MulFN-fl completely but not MulFN-~ (Bosveld et al, 1982;Vonk & Trapman, 1983).…”

Section: Gel Filtration Of Mulfn-~mentioning

confidence: 97%

“…Affinity chromatography over a monoclonal antiMuIFN-fl column (monoclonal antibody EB3) was performed as described previously (Vonk & Trapman, 1983). The flow-through fraction of this column was completely devoid of MuIFN-fl activity.…”

Section: Interferon Neutralization Assaymentioning

confidence: 99%

Isolation and Characterization of Subspecies of Murine Interferon Alpha

Lemson¹,

Vonk²,

Korput³

et al. 1984

Journal of General Virology

Self Cite

View full text Add to dashboard Cite

SUMMARYInterferon produced by mouse L-929 cells by incubation with poly(rI), poty(rC) is known to be composed of a mixture of MuIFN-c~ and MuIFN-fl. The c~ component was separated from the fl species by affinity chromatography over a monoclonal antiMuIFN-fl agarose column and partially purified by gel filtration. MuIFN-c~, prepared by this method was separated into at least five subspecies by chromatofocusing. The approximate pI values of these components are >~ 7.5, 6.5, 6.2, 5-9 and 5.6, respectively. Component 3 (pI 6.2) was the most prominent subspecies present in our MulFN-~ preparations, representing 40 to 50 ~o of the total antiviral activity. Component 1 (pI /> 7.5) which accounted for about 5~ of the antiviral activity on mouse cells, differed in some properties from the other interferon subspecies. It showed a relatively high antiviral activity on heterologous cells and it was eluted from a Sephadex column after the other c¢ subspecies. Furthermore, it showed a diminished binding to heparin as compared to the other MulFN-c¢ subspecies, indicating a lower affinity for polynucleotides.

show abstract

Large-Scale, One-Step Purification of Murine Interferon-Beta Using a Monoclonal Antibody

Cited by 8 publications

References 19 publications

Purification and carbohydrate structure of natural murine interferon‐β

Purification and carbohydrate structure of natural murine interferon‐β

Constitutive Expression of a Murine Interferon Alpha Gene in Hamster Cells and Characterization of Its Protein Product

Isolation and Characterization of Subspecies of Murine Interferon Alpha

Contact Info

Product

Resources

About