Purification and characterization of mouse interferon with novel affinity sorbents

Area of Science:

• Biochemistry
• Immunology
• Protein Chemistry

Background:

• Mouse interferon is a glycoprotein with therapeutic potential.
• Efficient purification methods are crucial for interferon characterization and application.
• Existing purification techniques may lack selectivity and efficiency.

Purpose of the Study:

• To develop novel selective sorbents for mouse interferon purification.
• To investigate the binding properties of interferon to different ligands.
• To enable efficient separation and characterization of mouse interferon components.

Main Methods:

• Immobilization of low-molecular-weight (hydrocarbons) and high-molecular-weight (bovine serum albumin) ligands on agarose.
• Chromatographic separation using selective sorbents.
• Analysis of interferon binding and recovery using varying ionic strengths.

Main Results:

• Selective binding of mouse L-cell interferon to immobilized ligands, primarily via hydrophobic interactions.
• A 2,000-fold increase in interferon specific activity (3 x 10^8 reference units/mg protein) in a single step with full recovery.
• 300-fold purification achieved using hydrocarbon-coated agarose.
• Identification of two major mouse interferon components through chromatography on bovine serum albumin-agarose, omega-carboxypentyl-agarose, and Bandeiraea simplicifolia lectin-agarose.

Conclusions:

• Novel sorbents effectively purify mouse interferon, significantly enhancing specific activity.
• Hydrophobic interactions play a key role in interferon binding to selective sorbents.
• This study provides the first efficient method for separating mouse interferon components, facilitating further research.