What question did this study set out to answer?

To establish a scalable method for producing high-titer mouse immune interferon (type II) and evaluate multiple chromatographic techniques for its partial purification.

January 1, 1979Infection and ImmunityOpen Access

Large-Scale Production and Partial Purification of Mouse Immune Interferon

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Authors

LOL C OsborneLabCorp (United States)JGJ GeorgiadesWashington University in St. Louis HJHoward M. JohnsonUniversity of Wisconsin–Madison

Discussion

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Overview

Laboratory study demonstrates high-yield purification of immune interferon from mouse spleen cells, revealing molecular weight heterogeneity.

Key Points

To establish a scalable method for producing high-titer mouse immune interferon (type II) and evaluate multiple chromatographic techniques for its partial purification.
Cultured mouse spleen cells in roller bottles stimulated with the T-cell mitogen staphylococcal enterotoxin A to generate interferon titers of 10^2.2 to 10^4 U/ml.
Concentrated crude harvests using 55% to 80% saturated ammonium sulfate precipitation.
Fractionated proteins sequentially using BSA-Affi-Gel 10 affinity chromatography, hydroxylapatite chromatography, and Ultrogel AcA 34 gel filtration.
Ammonium sulfate precipitation recovered 90% of interferon activity with a 20-fold concentration and a 2- to 6-fold purification.
BSA-Affi-Gel 10 chromatography achieved 100% activity recovery, yielding a 62-fold purification via pH gradient elution and a 390-fold purification via 1 M NaCl elution with specific activity reaching 2 x 10^5 U/mg.
Hydroxylapatite chromatography yielded 107-fold and 16-fold purifications across two fractions, while Ultrogel AcA 34 resolved two active peaks at molecular weights of ~40,000 and 70,000–90,000.

Cite This Study

Osborne et al. (1979) studied this question.

synapsesocial.com/papers/6a95b5203794e0e4a82bee37 https://doi.org/10.1128/iai.23.1.80-86.1979

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January 1, 1979Infection and ImmunityOpen Access

Large-Scale Production and Partial Purification of Mouse Immune Interferon

View Full Paper

Ask AI

Bookmark

Authors

LOL C OsborneLabCorp (United States)JGJ GeorgiadesWashington University in St. Louis HJHoward M. JohnsonUniversity of Wisconsin–Madison

Discussion

Member takes

Overview

Laboratory study demonstrates high-yield purification of immune interferon from mouse spleen cells, revealing molecular weight heterogeneity.

Key Points

To establish a scalable method for producing high-titer mouse immune interferon (type II) and evaluate multiple chromatographic techniques for its partial purification.
Cultured mouse spleen cells in roller bottles stimulated with the T-cell mitogen staphylococcal enterotoxin A to generate interferon titers of 10^2.2 to 10^4 U/ml.
Concentrated crude harvests using 55% to 80% saturated ammonium sulfate precipitation.
Fractionated proteins sequentially using BSA-Affi-Gel 10 affinity chromatography, hydroxylapatite chromatography, and Ultrogel AcA 34 gel filtration.
Ammonium sulfate precipitation recovered 90% of interferon activity with a 20-fold concentration and a 2- to 6-fold purification.
BSA-Affi-Gel 10 chromatography achieved 100% activity recovery, yielding a 62-fold purification via pH gradient elution and a 390-fold purification via 1 M NaCl elution with specific activity reaching 2 x 10^5 U/mg.
Hydroxylapatite chromatography yielded 107-fold and 16-fold purifications across two fractions, while Ultrogel AcA 34 resolved two active peaks at molecular weights of ~40,000 and 70,000–90,000.

Cite This Study

Osborne et al. (1979) studied this question.

synapsesocial.com/papers/6a95b5203794e0e4a82bee37 https://doi.org/10.1128/iai.23.1.80-86.1979

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