Large-scale production and partial purification of mouse immune interferon

Area of Science:

• Immunology
• Biochemistry
• Cell Biology

Background:

• Immune interferon (Type II) plays a crucial role in immune responses.
• Efficient large-scale production and purification methods are essential for studying its functions.
• Previous methods for interferon production and purification were limited in scale and efficiency.

Purpose of the Study:

• To establish a method for large-scale production of high-titered immune interferon.
• To develop purification strategies for immune interferon using various chromatographic techniques.
• To investigate the heterogeneity of purified immune interferon.

Main Methods:

• Large-scale production of immune interferon in roller cultures of mouse spleen cells stimulated with staphylococcal enterotoxin A.
• Concentration and partial purification using ammonium sulfate precipitation.
• Purification using affinity chromatography (BSA-Affi-Gel 10) and ion-exchange chromatography (hydroxylapatite).
• Analysis of molecular weight and heterogeneity using Ultrogel AcA 34 chromatography.

Main Results:

• Achieved high titers of immune interferon (10(2.2) to 10(4) U/ml).
• Ammonium sulfate precipitation resulted in 20-fold concentration and 2- to 6-fold purification.
• BSA-Affi-Gel 10 chromatography yielded up to 390-fold purification with specific activities of 2 x 10(5) U/mg protein.
• Hydroxylapatite chromatography achieved up to 107-fold purification.
• Ultrogel AcA 34 chromatography indicated heterogeneity with molecular weights of approximately 40,000 and 70,000-90,000.

Conclusions:

• A robust method for large-scale immune interferon production and purification was established.
• Chromatographic techniques, particularly BSA-Affi-Gel 10, are effective for purifying immune interferon.
• Immune interferon is a heterogeneous protein mixture with distinct molecular forms.