Preparation of the major plasma fractions by solid-phase polyelectrolytes

Area of Science:

• Biochemistry
• Polymer Chemistry
• Biotechnology

Background:

• Fractionation of plasma proteins is essential for therapeutic applications.
• Existing methods often involve harsh conditions or organic solvents, potentially denaturing proteins.
• There is a need for efficient, non-denaturing methods for isolating specific protein fractions.

Purpose of the Study:

• To develop a flexible, integrated batch system for isolating and purifying albumin, gamma globulin, and coagulation factors.
• To utilize solid-phase ethylene/maleic anhydride polymers for protein adsorption and elution.
• To establish non-denaturing methods for high-purity protein fraction recovery.

Main Methods:

• Utilized solid-phase ethylene/maleic anhydride (PE) polymers (E-5 and E-100).
• Employed a three-step batch process: adsorption, elution, and concentration (ultrafiltration/PEG).
• Relied on electrostatic bonding, avoiding organic solvents for a non-denaturing approach.

Main Results:

• Achieved high yields and purity for various fractions: coagulation factors (63% yield, 200-fold purification), albumin (92% yield, 98.5% purity), and gamma globulin (92% yield, 97.5% purity).
• Demonstrated successful isolation of antihemophilic factor (AHF) and von Willebrand factor (vWF).
• Observed removal of Hepatitis B surface antigen (HBsAg) and infectivity during gamma globulin preparation with E-100.

Conclusions:

• The developed batch method is flexible, efficient, and non-denaturing for plasma protein fractionation.
• Solid-phase PE polymers offer a versatile platform for isolating clinically important proteins.
• The method shows potential for simultaneously purifying therapeutic proteins and removing viral contaminants.