Methods for freezing and thawing proteins

Abstract

A method for freezing and thawing proteins is disclosed. The proteins are rapidly frozen by introducing droplets of a protein solution into a cryogenic freezing medium. The frozen pellets thus formed are rapidly thawed by introducing them into a heat transfer fluid to form a reconstitute protein solution.

Description

BACKGROUND OF THE INVENTION[0001]The invention provides for a method of uniformly freezing and thawing protein solution to minimize functional damage to the protein. More particularly, the invention provides non-equilibrium heat transfer to the protein solutions.[0002]Advances in biotechnology have led to increasing production of protein-based therapeutics. This in turn has led to increased demands for efficient methods to stabilize and store such therapeutic proteins. In a typical biomanufacturing system, the protein obtained from downstream processing and purification is usually in bulk quantity, in aqueous environment, and chemically stabilized by the addition of buffers and excipients. Sometimes additional preformulation studies are required following downstream processing for efficient stabilization. A protein solution that has been chemically stabilized often requires moderate to long periods of storage prior to final dosage formulation.[0003]Although lyophilization is one way...

AI Technical Summary

Benefits of technology

[0025]The cryogenic fluid that may be employed in the invention is selected from the group consisting of liquid nitrogen, oxygen, air and argon. The cryogenic fluid may not have to be subcooled should the effects resulting from gas formation not being an issue. The cryogenic fluid may be suitably processed such as by filtration processes to produce sterile fluids.

[0026]The cryogenic fluid may be maintained sub cooled by periodically subjecting to low pressure for short duration to cause partial boil off of cryogen.

Problems solved by technology

Although lyophilization is one way to improve stability of final drug products, it is not a practical or economical method for intermediate storage of protein solutions between bulk processing and final dosage formulation.

The secondary and tertiary structures are the ones that are susceptible to changes in microenvironments and ultimately cause the protein to change its conformation and lose functional activity.

This equilibrium unfolding of the protein during freezing is clearly a problem in terms of the storage and use of proteins.

Gaseous nitrogen, when formed, rises up as bubbles and meets downcoming droplets to cause turbulent contact at air / liquid interface which can damage proteins.

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