EFFECTIVE STABILIZATION OF VIRAL VECTORS IN LIQUID USING AN ALGORITHM-BASED DEVELOPMENT APPROACH
Author(s): ,
Eva Reinauer
Affiliations:
LEUKOCARE AG,Martinsried,Germany
,
Kristina Kemter
Affiliations:
LEUKOCARE AG,Martinsried,Germany
,
Julia Hasler
Affiliations:
LEUKOCARE AG,Martinsried,Germany
,
Carina Rodenstein
Affiliations:
LEUKOCARE AG,Martinsried,Germany
,
Jens Altrichter
Affiliations:
LEUKOCARE AG,Martinsried,Germany
Scholz Martin
Affiliations:
LEUKOCARE AG,Martinsried,Germany
EHA Library. Reinauer E. Jun 14, 2019; 266248; PF448
Eva Reinauer
Eva Reinauer
Contributions
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Abstract

Abstract: PF448

Type: Poster Presentation

Presentation during EHA24: On Friday, June 14, 2019 from 17:30 - 19:00

Location: Poster area

Background

With both the approvals of CD19 CAR-T cell therapies and promising clinical data of gene therapy trials such as in hemophilia or sickle cell disease, recent years have seen significant progress of gene and cell therapy in the field of hematological malignancies as well as rare hematologic genetic disorders. Hence, replication-deficient recombinant viral vectors such as adenovirus serotype 5, adeno-associated virus and members of the poxvirus family represent a rapidly growing field of vaccine development and gene therapy.

Viral vectors are known as complex supra-molecular ensembles of macromolecules produced by living organisms (nucleic acids, proteins, polysaccharides and in the case of lipid enveloped viruses of phospholipids) which are prone to a variety of complex chemical and physical degradation pathways, in particular due to stress induced by manufacturing, storage and distribution. This represents a significant hurdle for the development of stable vector-based pharmaceuticals such as vaccines or gene therapeutics

Aims

Here, we studied whether LEUKOCAREs formulation platform may also efficiently protect viral vectors in thermal stress experiments. As a representative example of viral vectors we used adenovirus serotype 5 (Ad5).

Methods

In an algorithm-based development approach, we used Design of Experiment (DoE) to select the most effectively stabilizing formulations for Ad5. We stored Ad5 at 37 °C and 25 °C as accelerated aging temperature to identify the most effective stabilizing excipients after short term storage as well as at 5 °C for real time storage. Based on these results, the best selected formulations were further iteratively optimized and used in long term storage at 5 °C.

Results
By analysis of the infectious virus titers and mathematical combination of these results with the DoE matrix, the linear influence of each amino acid used in the DoE matrix was determined. The accelerated aging conditions were shown to be predictive for real-time aging. Several of the excipients indicated a neutral influence, whereby we observed the importance of a well-balanced combination of the components with each other regarding concentration and stabilizing interactions. The predictive potential of this approach was confirmed by two iteratively composed  formulations tailored for Ad5 during long-term storage.

Conclusion
The pre-selection strategy of effectively stabilizing excipients by means of an algorithm-based development approach and the applied accelerated aging model is highly efficient and enables the generation of best-in-class stability formulations for Ad5 viruses and viral vectors in liquid. Moreover, this approach could have beneficial impact when applied early in downstream processing, which could enable significant reduction of viral vectors manufacturing costs for gene therapy.

Session topic: 24. Gene therapy, cellular immunotherapy and vaccination - Biology & Translational Res

Keyword(s): Cellular therapy

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