Purification of plasma protein

Area of Science:

• Biochemistry
• Protein Chemistry
• Hematology

Background:

• Coagulation proteins are present in plasma at trace levels.
• These proteins are susceptible to degradation and adsorption, complicating purification.
• Highly purified coagulation factors became available primarily after the 1970s.

Purpose of the Study:

• To review methods for purifying coagulation proteins from plasma.
• To highlight the role of chromatography, particularly affinity chromatography, in protein isolation.
• To discuss specific examples of successful purification strategies for various clotting factors.

Main Methods:

• Combination chromatography techniques including ion exchange, molecular sieve, and affinity chromatography.
• Utilizing specific adsorption properties, such as for Vitamin K dependent factors.
• Employing immobilized ligands in affinity chromatography, such as receptors, antibodies, substrates, or inhibitors.

Main Results:

• Affinity chromatography enables the isolation of specific proteins like thrombin (using benzamidine-Sepharose), factor VIII:C (using immunoadsorption), factor XI and prekallikrein (using high molecular weight kininogen), and antithrombin III (using heparin-Sepharose).
• Lysine-Sepharose affinity chromatography allows a single-step, high-yield purification of plasminogen from plasma, serum, or urine.
• These methods achieve the purification goals of high purity, biological activity, and high yield in simplified procedures.

Conclusions:

• Affinity chromatography is a powerful and versatile technique for purifying coagulation proteins.
• Specific ligand-protein interactions are key to effective and efficient protein isolation.
• The development of advanced chromatographic methods has significantly improved the availability of purified, active coagulation factors.