Periodic report I

Context and overall objectives of the project

What is the problem/issue being addressed in iConsensus?

Biopharmaceuticals are large molecules, most often proteins, used for therapy, e.g. antibodies. In contrast to small molecule drugs generated by organic synthesis, the biopharmaceuticals are produced using cells in well-regulated processes ('bioprocess') in bioreactor.

Today health technology is highly profitable but spending the highest percentage of its revenues on R&D. The development of a new drug costs about 1 billion dollars and takes ≥ 10 years. Reducing cost and time for the development of the manufacturing process represent very large savings and is essential to be first to market.

The worldwide community of bioprocessing is in quest for more rational tools to understand, predict and optimize the industrial biopharmaceutical manufacturing processes under development. To ensure the patient safety, these drug proteins are produced under very strict rules by the manufacturers and closely monitored by the Health Authorities.

Why is iConsensus important for society?

iConsensus aims to bring a disruptive new technology platform in mammalian-cell based production of biopharmaceuticals. It can be expected to reduce the time and costs for the process development and lead to a better manufacturing control. This improvement influences the time-to-market. Adding on for the biopharmaceutical European industry competiveness advantage, higher consistency, efficacy and robustness will be achieved. All these factors will generate reduced cost, failure and risk for product shortage.

The project clearly supports European Innovation, competiveness and industrial leadership while giving tools to improve the European well-being and societal challenges.

What are the overall objectives of iConsensus?

A cell culture used for the production of a biopharmaceutical is a complex mixture of components. Monitoring all the medium components (≥ 70) could provide a better knowledge of the process however it is a very large task. All these components can have an effect on the process performance, the purification process or on the product quality, which is the reason why monitoring all these compounds is advantageous.

To address the issues listed above, iConsensus will provide innovative analytical, hardware, software and high-throughput (HTP) solutions for the development, monitoring and control of mammalian cell cultivation process producing biopharmaceuticals. iConsensus will develop an analytical platform with on-line/at-line sensors and detection methods measuring multiple factors in the cell culture and the produced therapeutic protein with monoclonal antibody taken as one of the models. Under advanced data management, the analytical platform will be integrated in new micro-bioreactors for HTP screening on one hand and in classical stirred tank vessel bioreactors on the other hand. Based on the analytical information, feedback control will be applied. Implementation in GMP production of the sensing and control tools will be a major goal.

Main results achieved so far

High-throughput microbioreactors (MBR)

In iConsensus, novel microbioreactors for mammalian cell culture development are created. Designing and characterization of the reactor’s geometry have been completed according to the requirements for mammalian cell culture. The regulations of oxygen concentration and pH of these new MBR are highly satisfactory.

CE chip platform

This work focuses on developing methods based on capillary electrophoresis (CE) to measure different components in the culture and to transfer these into chip format. The transfer of preliminary methods to CE chip setups has been successful for three types of metabolites.

Optical on-line sensors

Several sensors based on optical properties are created iand proof-of-concept's have been demonstrated for several of these. New formats have also been created for pH, oxygen and CO2 sensors for different bioreactors and vessels, including autoclavable sensors.

Spectrometry and chemometry support

In WP5, a generic chemometric PLS regression prediction model was created to predict glucose, glutamate and lactic acid concentration with errors of less than 5 % by using Raman spectra from different CHO cultivations issued by different Raman spectrometer.

Holographic methods

Iprasense has developed a new concept for the quantification of the cell concentration based on holographic image. The evaluation carried out at GSK indicated very close correlation (<10% error) with a reference instrument. In addition, the high refractive index population is correlated to early detection of viability drop, and a correlation to early apoptotic cells has been shown.

Affinity-based microfluidics

Russom's team at KTH has developed a multiplexed bead-based microfluidic device to perform four immunoassays targeting three different analytes and a control on a single sample. Colorimetric quantification of CHO host cell proteins as key impurities, IgG as target biopharmaceutical and LDH as a marker of cell viability was performed directly from spiked cell culture medium and limits of detection in the low ng/mL range.

Analysis of glycosylation

PAIABio has continued the validation of its lectin based detection technology with samples from Bayer on different antibody glycoforms. Simplified glycan assay workflow has been created with KTH to facilitate the transfer into microfluidics. First results from samples of Byondis show that this approach is feasible.

Total integration and data management, MID

In iConsensus, an integrated analytical system will created, 'At-line integrated analytical system' or 'ALIAS'. The first year the architecture and functionalities of the integrated system have been defined and in the second year fine-tuned, together with the monitoring information database (MID).

Modeling and control of animal cell cultures

To support the modelling, KTH has performed experiments with information on the main metabolites including the amino acids. Mechanistic kinetic model by column generation and statistical approach for the kinetics as well as data-driven models have been generated and will be improved in the next period of the project. The first results are promising and show the capacity to predict the behaviour of the process, for instance to improve the medium formulation.

Progress beyond the state of the art, expected results until the end of the project and potential impacts

iConsensus aims at developing a new platform for real time analysis of the components. Nowadays, the development of cultivation processes uses empirical methods supported by statistics, and high-throughput, where only very few parameters are easily quantified, leading to suboptimal outcome. When more efforts are put on the analytical technology, this requires enormous resources and time. To address this, iConsensus brings solutions by far pioneer, with no equivalent product today, with real-time solutions and advanced MBR. Besides the importance of the use of innovative tools in development it is also crucial to have reliable, fast and qualified analytical tools for commercial manufacturing processes. Data and knowledge gained during development should be usable in a straight line in the final manufacturing process. The possibility to use the same analytical tools throughout the development and life cycle management of a process/product will be highly beneficial for the control and robustness of the final manufacturing process (Process Analytical tools, Design Space).