Microassay for Human and Chick Cell Interferon

Tilles, Jeremiah G.; Finland, Maxwell

doi:10.1128/am.16.11.1706-1707.1968

Cited by 32 publications

(1 citation statement)

References 2 publications

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“…Interferon Assay. The interferon end point was determined by a cytopathic effect inhibition assay (Tilles & Finland, 1968; Armstrong, 1971). Samples were assayed in 96-well microtitration plates (Falcon, Oxnard, CA).…”

Section: Methodsmentioning

confidence: 99%

Purification of human .gamma.-interferon to essential homogeneity and its biochemical characterization

Braude¹

1984

Biochemistry

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A multistep procedure has been developed which enables human gamma-interferon (HuIFN-gamma) to be purified to essential homogeneity. The procedure takes advantage of a modification of a previously described sequential chromatographic technique [Braude, I.A. (1983) Prep. Biochem. 13, 177-190] and the high isoelectric point of HuIFN-gamma (pH 9.5-9.8). The steps include Controlled Pore Glass adsorption chromatography, concanavalin A-Sepharose and heparin-Sepharose affinity chromatography, cation-exchange chromatography, and gel filtration chromatography. The purified HuIFN-gamma had a specific activity of 5.9 X 10(7) units/mg. This represents a purification of more than 70 000-fold and a 33% recovery. In addition, one gel filtration fraction had a specific activity of 2.5 X 10(8) units/mg. This represents a purification of greater than 300 000-fold and a recovery of greater than 17%. This fraction, when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was shown to be composed of one major 26-kilodalton (kDa) species and four minor species of 74, 67, 56, and 22 kDa. Analysis of this material with anti-HuIFN-gamma monoclonal antibody immunoabsorbent columns indicates that both the 26- and the 22-kDa species are HuIFN-gamma. Thus, the final product is essentially homogeneous (90-92% HuIFN-gamma), and the specific activity of pure HuIFN-gamma is approximately (2.7-2.8) X 10(8) units/mg of protein. Finally, the 26- and 22-kDa moieties are shown to be similar, if not identical, proteins as judged by amino acid and sequence analyses.

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Section: Methodsmentioning

confidence: 99%

Purification of human .gamma.-interferon to essential homogeneity and its biochemical characterization

Braude¹

1984

Biochemistry

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Inhibition of leukemic cell proliferation by factor(s) released from irradiated lymphocytes of B‐chronic lymphocytic leukemia patients

et al. 1994

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An attempt was made to clarify the mechanism by which splenic irradiation in patients with B-chronic lymphocytic leukemia (B-CLL) can induce a reduction in lymph node size. For this purpose peripheral blood lymphocytes from B-CLL patients were exposed to cobalt irradiation and were cultured for 1-8 days. The effect of the supernatants on the proliferation capacity of normal and malignant human cells was examined. A suppression of phytohemagglutinin (PHA)-induced proliferation of autologous and heterologous B-CLL lymphocytes was observed, whereas there was no effect on the proliferation of lymphocytes obtained from healthy volunteers. In addition, supernatants of irradiated B-CLL lymphocytes inhibited thymidine incorporation into blasts derived from patients with acute leukemia and the lymphoblastoid cell line Daudi, but they did not exert any effect on normal cells obtained from human embryonic liver. These results suggest the secretion of some factor(s) by irradiated B-CLL lymphocytes, which may inhibit the proliferation of malignant cells but has no effect on normal cells.

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Assay of Interferons

Grossberg¹,

Jameson²,

Sedmak³

1984

Interferons and Their Applications

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Microassay for Human and Chick Cell Interferon

Abstract: The microassay for human and chick interferon described in this paper required much smaller amounts of samples and reagents and considerably less time and effort than the plaque reduction assay while yielding comparable results.

Cited by 32 publications

References 2 publications

Purification of human .gamma.-interferon to essential homogeneity and its biochemical characterization

Purification of human .gamma.-interferon to essential homogeneity and its biochemical characterization

Inhibition of leukemic cell proliferation by factor(s) released from irradiated lymphocytes of B‐chronic lymphocytic leukemia patients

Assay of Interferons

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