Event: Cell and Gene Therapy Insights webinar (February 17, 2022)
Presenter: Ales Strancar, Managing Director
Title of the lecture: AAV manufacturing process development using fast USP and DSP biochromatography analytics
Key learning points:
- One of the key challenges in manufacturing viral vectors is to increase the ratio between empty and full capsids.
- When the expression in the cell line results in less than 10% full capsid it is quite impossible to reach better than 90% full in the final product. It is therefore mandatory to optimise the USP to result in better empty/full ratio.
- This can be efficiently realised by using at-line PATfix to allow for analysis of the full and empty capsids ratio directly in the harvest.
- The residual empty capsids can be removed by polishing step using different anion exchange columns.
GET in TOUCH:
Webpage: [ Ссылка ]
E-mail: sales@biaseparations.com
Tech support: help@biaseparations.com
CHAPTERS:
0:00 Introduction
2:50 Experts in express bioproduct manufacturing process development -PATfix enables the speed
7:05 Rapid chromatography processes by convective mass transport
11:40 PATfix components: build-in methods and dedicated columns
13:09 CIMac analytical columns for in-process HPLC - no carry over of contaminants or viruses
14:07 To enable fast process development high performance analytics is mandatory PATIE HPLC with multiple detectors allows for sample characterisation in an hour.
18:13 Orthogonal tools to study AAV particles distribution - HPLC: ultracentrifuge
22:00 PATfix two columns switching method to analyze crude AAV samples
26:09 Case study. Transfection reagent number 3 provided best AAV titre and E/F ratio
29:02 Intracellular produced AAV - after cell lysis sample is reach with Chromatin
32:37 Chromatin removal options
34:42 Example: Chromatin Solid phase extraction (SPE) using particles
38:01 Influence of Mgon separation of AAV capsids using CIM QA column
41:55 Density gradient ultracentrifugation to check the purity of full capsids
43:49 Conclusions - Ultracentrifugation and chromatography support orthogonal analysis. Combined
47:24 WEBINARS CELL & GENE THERAPY INSIGHTS ANV manufacturing process development using fact USP and DSP biochromatography analysis
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