An Advanced Monitoring Platform for Rational Design of Recombinant Processes - Measurement, Monitoring, Modelling, and Control of Bioprocesses - page 66

OUR Oxygen uptake rate

ppmV Parts per million volume

pptV Parts per trillion volume

PLS Partial least squares

PCN Plasmid copy number

PTR-MS Proton transfer reaction mass spectrometry

QbD Quality by design

RBF-NN Radial basis function neural network

RT-PCR Real-time polymerase chain reaction

RQ Respiration quotient

RMSEP Root-mean-square error of prediction

SOD Superoxide dismutase

TCN Total cell number

FDA US food and drug administration

VOC Volatile organic compound

Contents

1 Introduction.......................................................................................................................... 66

1.1 Bioprocess Monitoring ............................................................................................... 67

1.2 Recombinant Protein Expression in E. coli Systems ................................................ 69

2 A Process Monitoring Platform for Recombinant Protein Production.............................. 70

2.1 Monitoring of Biomass and Growth .......................................................................... 70

2.2 Monitoring of Metabolites and Cell Compounds...................................................... 71

2.3 Monitoring of Products from the Recombinant System ........................................... 72

2.4 Monitoring of Cellular Response to Recombinant Gene Expression....................... 74

3 Chemometry and Prediction................................................................................................ 76

4 Process and Systems Design............................................................................................... 77

4.1 Transcription Tuning .................................................................................................. 78

4.2 Stabilized Plasmid Copy Number .............................................................................. 79

5 Plasmid-Free Systems ......................................................................................................... 80

6 Conclusions.......................................................................................................................... 81

References.................................................................................................................................. 81

1 Introduction

Recombinant biopharmaceutical production processes require qualified process and
systems understanding for rational host and process design. As in other industrial
processes, yield and product quality are the most important targets in biotechno-
logical production. Due to tight regulatory requirements for manufacturing of