I1 - Vector Product Engineering, Development and Manufacturing (excluding AAV)
1858: Novel Ligand Outperforms State-of-the-Art Ligand in Lentivirus Purification by Anion Exchange Chromatography
Type: Poster Session
Poster Board Number: 1858
Presentation Details
Session Title: Friday Posters: Vector Product Engineering, Development, and Manufacturing
Lentivirus vectors (LVV) are widely used in cell engineering for cell therapy applications, and are leveraged in the manufacturing of an increasing number of products such as CAR-T cells. LVV used for cell engineering are produced using cell expression systems at varying scales and thus their isolation from conditioned media containing host-cell and cell culture-derived contaminants is a crucial step during production. LVV purification is usually performed by anion exchange chromatography (AEX), but the state-of-the-art run conditions required often lead to significant loss of functional virus due to LVV’s high sensitivity to pH, salt concentration, and shear stress.
Here we present a novel functionalized resin that enhances the purification of LVVs, enabling the optimized high yield recovery of functional LVV under mild conditions, outperforming the benchmark ligand used for LVV purification. Different resin ligands were screened for optimal binding and elution of enveloped biological particles, with optimal elution conditions at salt concentrations ranging between 150-275 mM and pH 7.0-8.0. Collected fractions were analyzed for viral recovery by HEK293T cell transduction assay and p24 ELISA. Depletion of contaminants was assessed by total protein quantification assay, HEK293 Host Cell protein ELISA and QuantiFluor dsDNA assays.
We show that LVV can be successfully purified in both capturing and polishing mode from a diverse set of clarified conditioned media. Due to the mild elution conditions, viral titer recoveries of up to 4-fold higher were achieved with nearly 2-fold improved contaminants clearance (HCP and dsDNA) compared to the benchmark DEAE ligand. Our novel ligand is a promising alternative in LVV downstream processing, leading to significant increases in process productivity by recovering purer LVV at high yield.
Nicolas Meier, Andreia M. Silva, Fabian Trick, Joel de Beer
Anjarium Biosciences AG, Schlieren, Switzerland"
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