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Upstream processes are rather complex to design and the productivity of cells under suitable cultivation conditions is hard to predict. The method of choice for examining the design space is to execute high‐throughput cultivation screenings in micro‐scale format. Various predictive in silico models have been developed for many downstream processes, leading to a reduction of time and material costs. This paper presents a combined optimization approach based on high‐throughput micro‐scale cultivation experiments and chromatography modeling. The overall optimized system must not necessarily be the one with highest product titers, but the one resulting in an overall superior process performance in up‐ and downstream.
The methodology is presented in a case study for the Cherry‐tagged enzyme Glutathione‐S‐Transferase from Escherichia coli SE1. The Cherry‐Tag(tm) (Delphi Genetics, Belgium) which can be fused to any target protein allows for direct product analytics by simple VIS absorption measurements. High‐throughput cultivations were carried out in a 48‐well format in a BioLector micro‐scale cultivation system (m2p‐Labs, Germany). The downstream process optimization for a set of randomly picked upstream conditions producing high yields was performed in silico using a chromatography modeling software developed in‐house (ChromX). The suggested in silico‐optimized operational modes for product capturing were validated subsequently. The overall best system was chosen based on a combination of excellent up‐ and downstream performance. This article is protected by copyright. All rights reserved
Abstract The single radial immunodiffusion assay has been the accepted method for determining the potency of inactivated influenza vaccines since 1978. The world‐wide adoption of this assay for vaccine standardisation was facilitated through collaborative studies that demonstrated a high ... mehr
Abstract Background Whether morbidity from the 1918‐19 influenza pandemic discriminated by socioeconomic status has remained a subject of debate for 100 years. In lack of data to study this issue recent literature have hypothesized that morbidity was “socially neutral”. Objectives ... mehr