Work package 4

Project Title: Disposable paper-based immunosensor biochips for rapid monitoring of recombinant protein purity in up- and downstream processing (WP4)

ESR 5    

Host: Acreo, Sweden

Objectives: To develop a new versatile real-time biochip for rapid and easy measurement of protein product forms that facilitate process R&D as well as plant operation. The printed electronics technology should allow low-cost analytical methods (single use strips in paper materials) which could be used in typical R&D and manufacture environments. The biological recognition elements (antibodies, fragments) will be interactively with the other partners (SAN, FUJI) related to target drugs, e.g. a recombinant protein and its adverse forms. Recognition elements and complementary reagents will be deposited and immobilised in the printed chip. Lateral diffusion techniques will be considered if necessary to apply the target samples. An essential part of the methodology work is to establish and accomplish robust and versatile electronics between the immune-active recognition elements and the user interface

Expected Results: Prototype of a printed imunosensor biochip for product measurement will be manufactured and tested in industrial conditions.

 

 

Project Title: Disposable paper-based biochips for rapid culture media evaluation (WP4)

ESR 6    

Host: Acreo, Sweden

Objectives: To develop a new at-line real-time biochip capable of simultaneous monitoring of key media components in cultivation media. The at-line device will support process development and have potential of in-process monitoring at a manufacturing plant. Use of printed electronics technology should allow maintenance free monitoring that significantly facilitates implementation of new measurement methods. Biological recognition elements suitable for media components detection, e.g. enzyme based assays, ELISAs, as well as spectrophotometry assay reactions for key compounds (e.g. glucose, amino acids, overflow metabolites etc.) will be identified in conjunctions with ITN partners SAN, FUJI and CHN. Assay reagents will be deposed in the printed chip. Lateral diffusion techniques will be used to apply the target samples and diffuse assay reagents. The assay reaction will be detected by electronic or optical sensing. Focus of efforts will be on the biomolecular and bioanalytical solutions.

Expected Results: Prototype of a printed media sensor biochip will be manufactured and tested in industrial conditions.

 

 

Project Title: Real-time monitoring by soft sensors for quality parameters in recombinant protein production (WP4)

ESR 11  

Host: Linkoping University, Sweden

Objectives: To develop an on-line sensor solution where hardware sensors are combined with a mathematical model for the bioprocess and where the critical quality attributes (CQAs) are the analytical targets. The CQAs are selected so they are relevant in a recombinant protein process. The hardware part of the sensor is based on solutions developed in project 4.A and 4.B. The soft sensor technology at LIU will be used with existing bioprocess instrumentation data (e.g. electrodes, flow meters, optical detectors, pressure and temperature sensors) as inputs into mechanistic or multivariate models. In a bioprocess this must be supported by additional input data from on-line sensors. The ESR will apply such data from optical (fluorescence, infrared, imaging) and electronic sensor chips (printed electronics from 4.A and 4.B) with mathematical models. Focus for the ESR will be on building and evaluating the soft sensor models using cases close to the other partners’ actual systems. The deliverable will be two soft sensors adapted to fed-batch recombinant production and agreed CQAs.

Expected Results: This project will provide an in-depth evaluation of modelling methods suitable for soft sensor development for the industrial case studies used in this ITN. The resulting soft sensor solution will be tested on industrial case study systems and will form a crucial part of the resulting rapid bioprocess development framework.

 

 

Project Title: Real-time monitoring by soft sensors for quality parameters in downstream processing (WP4)

ESR 12  

Host: Linkoping University, Sweden

Objectives: To develop an at-line sensor for detection of impurities downstream where hardware sensors are combined with a mathematical model for the bioprocess and where CQAs are analytical targets. The CQAs are typical protein product forms occurring in recombinant protein processing. The hardware part of the sensor is based on solutions developed in project 4.A.  The methodological principle described in 4.C above will be pursued in this project - soft sensors using existing bioprocess instrumentation (e.g. electrodes, flow meters, optical detectors, pressure and temperature sensors) as inputs into mechanistic or multivariate models. For downstream processes (filtration units, centrifuges, chromatography columns) monitoring is carried out predominantly at the units’ effluents and control by adjusting e.g. flow rate, elution procedures, and speed rates. On-line/at-line monitoring with optical (e.g. NIR, fluorescence, turbidity) or electric sensors (e.g. conductivity, capacitance) provides signals for software based modelling. The model estimates of impurities can be used as inputs to control the processes. The ESR will investigate these possibilities and develop a soft sensor specifically for this purpose.

Expected Results: This project will review the most appropriate methodology for soft sensors in downstream processing and develop two soft sensor solution for chromatographic protein purification, which will be tested on industrial case studies.

Project Title: Exploiting integrated rapid bioprocess monitoring methods for acceleration of process development (WP4)

ESR 3    

Host: Newcastle University, UK

Objectives: Integrate the analytical and modelling methods developed in all WPs into a framework for accelerated process development. Methods to build process model toolbox need to address knowledge diversity in addition to specifying process model structure and capability and the means by which the process model is parameterised. Among the important innovations is the ability of such models to incorporate a quantification of belief that can be utilised to determine the risk. Bayesian strategies offer both this capability and the opportunity to refine the representation as knowledge is accumulated. Another representational challenge is the ability to capture the behaviour of a whole process rather than individual unit operations. The difficulty involves the capture of unit to unit interactions and the representational requirements of other units to build a whole process description. Here agent based strategies offer great potential. Foremost among all the representational problems faced is the need to specify structure. Hybrid relationships offer a means of using mechanistic knowledge but other approaches can be used to capture structure, evolutionary computational approaches offer a practical alternative.

Expected Results: This project will review the most appropriate methodology for general monitoring framework in bioprocessing and develop the overall rapid bioprocess development framework integrating all the model representations from the remaining BioRapid projects.