Monoclonal antibodies to human immune interferon and their application for affinity chromatography

Le, Junming; Barrowclough, Barbara S.; Vilček, Ján

doi:10.1016/0022-1759(84)90277-1

Cited by 40 publications

(14 citation statements)

References 17 publications

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“…Elution with SDS-containing buffer and subsequent SDS-PAGE analysis of the eluate yields several bands in the 20-to 25-kd range, which are not found to be eluted when conditioned media lacking IFN-y arc applied to this column. This is consistent with the reported molecular weight of HuIFN-y observed on SDS-PAGE [3,14], The development of several other mAb to HuIFN-y have been recently reported [5][6][7][8][9], Hochkeppel and de Ley [5] isolated a mono clonal antibody, identified as being specific for HuIFN-y on the basis of its ability to pre cipitate a 46 kd molecule from a crude IFN-y preparation. Oleszak et al [9] developed an IgM mAb specific for HuIFN-y.…”

Section: Discussionsupporting

confidence: 74%

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A High-Affinity Monoclonal Antibody (GIF-1) to Human Gamma-interferon: Neutralization of Interferon Mediated Inhibition of Retrovirus Production and 2’-5’ (A) Synthetase Induction

et al. 1986

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An hybridoma clone secreting an IgG 1 monoclonal antibody (GIF-1) specific for human gamma-interferon (HuIFN-γ) has been generated using HAT medium supplemented with insulin (HIAT) at the initial stage of cell fusion. This antibody is capable of neutralizing the antiviral activity of HuIFN-γ, the ability of HuIFN-γ to inhibit retroviral replication in RD-114 cells, and the ability of HuIFN-γ to induce the 2’-5’ oligoadenylate (A) synthetase in RD-114 and HeLa cells. Eluate from an immunoaffinity column containing GIF-1 yielded two protein bands of molecular weight of 20 and 25 kd when subjected to SDS-PAGE.

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Section: Discussionsupporting

confidence: 74%

“…Immunoaffinity chromatog raphy gave a 50-fold purification of crude IFN-y. Le et al [6,7] have reported on 2 mAb specific for HuIFN-y. These are both of the IgG| subclass.…”

Section: Discussionmentioning

confidence: 99%

A High-Affinity Monoclonal Antibody (GIF-1) to Human Gamma-interferon: Neutralization of Interferon Mediated Inhibition of Retrovirus Production and 2’-5’ (A) Synthetase Induction

et al. 1986

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“…The IRMA described in this paper should facilitate work with both natural and recombinant E. coli-derived human IFN-y. Unlike monoclonal antibody GIF-1 (31), which recognizes only natural and not E. coli-derived recombinant IFN-y (14,19), both antibodies B1 and B3 react equally well with glycosylated natural and unglycosylated E. coli-derived human IFN-y (18,19). Since the IRMA can detect as little as 0.1 NIH reference unit/ml of IFN-y, the IRMA is more sensitive than the common biological assay in which the limit of detectability usually is around 4 units/ml.…”

Section: Resultsmentioning

confidence: 99%

“…Recently, two mouse monoclonal antibodies, B1 and B3, specific for natural and Escherichia coli-derived recombinant human IFN-y have been developed (18,19). The two antibodies recognize different epitopes, since B1 and B3 differ in their reactivity with the IFN-y molecule (18) and they do not compete with each other's binding to IFN-y (unpublished data).…”

mentioning

confidence: 99%

Use of monoclonal antibodies as sensitive and specific probes for biologically active human gamma-interferon.

Chang¹,

McKinney²,

Liu³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

106

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Mouse monoclonal antibodies Bi and B3 are specific for natural and Escherichia coli-derived recombinant human V-interferon (IFN-y). The two antibodies recognize different epitopes of the IFN-y molecule and do not compete with each other's binding. We have used these two antibodies to construct a solid-phase, sandwich immunoradiometric assay for human IFN-y. Purified antibody Bi was coated on polystyrene beads (0.64 cm in diameter) and used as the solid-phase immunoadsorbent and antibody B3 was labeled with 1251 and used as tracer. This assay can be completed in about 4 hr and is capable of detecting IFN-y levels in human serum or tissue culture fluids as low as 0.1 NIH reference unit/mi. Recombinant human IFN-y derived from E. coli was detectable at a concentration of 0.02 ng/ml. The assay appears to be specific for the biologically active forms of IFN-y, since after exposure to pH 2, 370C, or 560C, biological activity and reactivity in the immunoradiometric assay decreased in parallel. The immunoradiometric assay can be employed for the analysis of the structural characteristics of the human IFN-y molecule.y-Interferon (IFN-y), also called immune IFN, is a lymphokine secreted by T lymphocytes upon stimulation with antigens or nonspecific mitogens (1)(2)(3)(4)(5)(6). Like IFN-a and -3(7-9), IFN-y induces the resistance of cells to viral infection and also exerts a range of immunoregulatory activities in vivo and in vitro, such as the augmentation of natural killer cell tumoricidal activity (4). In addition, IFN-y activates macrophages by triggering their morphological transformation, secretion of soluble factors, and expression of surface Ia antigens and Fc receptors (10, 11). Recent evidence also strongly suggests that IFN-y is indistinguishable from macrophage activation factor (12-16).Routine laboratory assays for IFNs are generally based on their ability to inhibit the lysis of cultured cells by viruses (17). Recently, two mouse monoclonal antibodies, B1 and B3, specific for natural and Escherichia coli-derived recombinant human IFN-y have been developed (18, 19). The two antibodies recognize different epitopes, since B1 and B3 differ in their reactivity with the IFN-y molecule (18) and they do not compete with each other's binding to IFN-y (unpublished data). Using these two antibodies, we have developed a sensitive and rapid solid-phase immunoradiometric assay (IRMA) for IFN-y. Our report shows that this assay can be used as a specific probe for biologically active human IFN-y. MATERIAL AND METHODSPreparation of IFN-y and Sources of Other IFNs. Human IFN-y was produced in the cultures of lymphocyte-rich plateletpheresis residue induced with phytohemagglutinin and phorbol 12-myristate 13-acetate. It was partially purified by a four-step protocol (20) Preparation of Bi Antibody-Coated Polystyrene Beads. Polystyrene beads, 0.64 cm in diameter (Precision Plastic Balls, Chicago), were washed with ethanol and phosphatebuffered saline (Pi/NaCl, pH 7.0). Coating of the beads with B1 antibody was performed...

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“…The monoclonal antibody B3 [25] against purified nIFN-y was a gift of Dr. J. Vilcek, New York University, New York. The monoclonal antibody M 0 2 was a gift of Takeda, Osaka, Japan.…”

Section: Purified Antibodies To Ifnmentioning

confidence: 99%

Identity between human interferon‐γ and “macrophage‐activating factor” produced by human T lymphocytes

et al. 1986

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Human peripheral blood monocytes purified by counterflow elutriation were activated in vitro by human natural or recombinant interferon-gamma (IFN-gamma) as shown by enhanced killing of Listeria monocytogenes and increased production of H2O2 in response to phorbol myristate acetate. Half-maximal stimulation for macrophage activation (MAF) was observed with 10-20 antiviral U/ml of purified recombinant IFN-gamma. These MAF activities were found to correlate with the antiviral activity dependent on IFN-gamma under several experimental conditions. Both activities were recovered together from supernatants of concanavalin A-stimulated peripheral blood mononuclear cells and in the media of a large number of T cell clones of different specificities. The parallelism between the two activities was also observed upon fractionation of culture media from producing cells and upon treatment of such preparations with low pH and high temperature. Finally, three antibodies with different specificities were found to abrogate the MAF and antiviral activities from lymphocyte culture supernatant. These results indicate that MAF released by stimulated lymphocytes is identical to IFN-gamma.

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Monoclonal antibodies to human immune interferon and their application for affinity chromatography

Cited by 40 publications

References 17 publications

A High-Affinity Monoclonal Antibody (GIF-1) to Human Gamma-interferon: Neutralization of Interferon Mediated Inhibition of Retrovirus Production and 2’-5’ (A) Synthetase Induction

A High-Affinity Monoclonal Antibody (GIF-1) to Human Gamma-interferon: Neutralization of Interferon Mediated Inhibition of Retrovirus Production and 2’-5’ (A) Synthetase Induction

Use of monoclonal antibodies as sensitive and specific probes for biologically active human gamma-interferon.

Identity between human interferon‐γ and “macrophage‐activating factor” produced by human T lymphocytes

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