Immobilized cobalt affinity chromatography provides a novel, efficient method for herpes simplex virus type 1 gene...

Area of Science:

• Gene therapy vector development
• Virology and molecular biology
• Bioprocessing and purification technologies

Background:

• Herpes simplex virus type 1 (HSV-1) shows promise as a gene therapy vector, especially for peripheral nerve applications.
• Scalable, high-yield manufacturing of pure gene vectors is crucial for clinical trials.
• Enveloped viruses like HSV-1 necessitate gentle purification methods, avoiding centrifugation and cellular contaminants.

Purpose of the Study:

• To develop a novel, gentle, and scalable purification method for HSV-1 gene therapy vectors.
• To engineer HSV-1 to display a specific tag for selective purification.
• To assess the efficacy of immobilized metal affinity chromatography (IMAC) using cobalt for HSV-1 purification.

Main Methods:

• Engineered HSV-1 to display a cobalt-binding peptide in the glycoprotein B envelope domain.
• Developed an immobilized metal affinity chromatography (IMAC) method utilizing cobalt ions.
• Optimized IMAC loading conditions and minimized cobalt ion leakage.

Main Results:

• Recovered 78% of the engineered HSV-1 recombinant virus.
• Achieved a >96% reduction in contaminating cellular proteins and DNA.
• Cobalt-based IMAC demonstrated selective binding to the tagged HSV-1 without significant interaction with normal viral components or contaminants.

Conclusions:

• Cobalt-dependent IMAC is a highly effective and selective method for purifying HSV-1 gene therapy vectors.
• This novel purification strategy offers a scalable and gentle approach, crucial for producing clinical-grade viral vectors.
• The engineered HSV-1 with a cobalt-binding tag provides a robust platform for advanced gene therapy manufacturing.