Industrial Placement Student FUJIFILM Diosynth Biotechnologies Ellie Hawke Industrial Placement Student FUJIFILM Diosynth Biotechnologies Confidential

Presentation overview Background My placement experience An introduction to FUJIFILM Diosynth My experiences at FUJIFILM Diosynth Advantages of an industrial placement

Background Initially applied to BSc in Biochemistry. Transferred onto Biomedical Sciences at the end of my first year. Currently undertaking an Industrial Placement Year at Fujifilm Diosynth. As a result of my placement year I now have aspirations to complete a PhD and return to industry. Confidential

My placement experience Up until the early part of second year I had no aspirations to undertake an industrial placement year. Attended placement talks and started to prepare CV. Application process Interview Start

An introduction to Fujifilm Diosynth. cGMP Contract Development and Manufacturing Organisation (CDMO) Based in Billingham, Wilton, North Carolina and Texas. Various departments including : - Protein expression- E.coli, Yeast and CHO capabilities. - Downstream purification - Analytical services - Large scale manufacturing Confidential

My experiences at Fujifilm Diosynth – an introduction to Biologics Utilisation of the internal cell machinery to manufacture drugs. More complex drugs require different expression systems. No post translational modifications required; E. coli could be used. If post translational modifications are required, such as glycosylation, a mammalian expression system would be used. In Mammalian Cell Culture we are producing monoclonal antibodies using a CHO expression system. Apollo™’ – a standard platform process we follow, producing a stable cell line expressing the desired antibody. Confidential

Apollo™ CLD process 1) Transfection & selection Vector 1) Transfection & selection 2) First screening & cloning step: ClonePix™2 Clone-27 5) Expansion, second 96-well plate screen & second shaken, fed-batch multi-well plate screen 3) Expansion, first 96-well plate screen & first shaken, fed-batch multi-well plate screen 4) Animal component free limiting dilution cloning 8a) Cell line stability study 6) ambr15™ Screen 7) Research cell bank 8b) Cell banking activities Confidential Say this is a look at the entire Apollo process – don’t worry if you don’t understand. Main points, start with inserting DNA into the host cell line that tells the cells which protein to make. Screening steps as we want to choose the cells that produce the highest amount of proteins. About 18weeks long. Research cell bank at the end – make frozen vials of top producing cell lines and store in liquid nitrogen, used for cell banking which goes into GMP manufacturing - Large volumes, may be needed for clinical trials or toxicity studies etc

Experience – The Cytomine. Sphere Fluidics Cyto-Mine® Technology. Based on a FRET-assay which allows the sorting of picodroplets. The cytomine is a high-throughput instrument that uses picodroplet technology and microfluidics to process up to 40 million heterogeneous mammalian cells in less than a day. Confidential https://www.youtube.com/watch?time_continue=46&v=N9XpZHvnzys

Experience Currently the 18 week timeline includes 2 cloning steps but with the implementation of the cytomine CLD timelines can be reduced to 10 weeks. Whilst working at Fujifilm I have been working to try and validate the cytomine machine as a monoclonality assurance method so that it can be used in CLD process This ultimately speeds up the delivery of customer projects without compromising product quality. Confidential 18 week timeline is Apollo platform but with the implementation of the cytomine the new improved platform will be launched around April time.

CytoMine Worflow 5) Single droplet dispensing Selected picodroplets are dispensed into a well of a 96 well plate. 5 images are taken pre dispensing and a heat map of the 96 well plates is produced. 1)Sample Loading The prepared cell suspension and assay reagents are loaded into a Cytocartridge and the Cytocartridge is loaded into the machine. 6) 96 well plates are imaged at set time points using the CellMetric™. Colony formation can be tracked back to the single cell stage. 2) Picodroplet generation Cells and assay reagents are encapsulated into picodroplets. Hundreds of thousands of cells can be encapsulated and processed in a single run. 3) Productivity screen Picodroplets are incubated. Cells secrete antibody product and due to encapsulation, the product produced by a single cell can be detected. 4)Droplet sorting Picodroplets which have the highest productivity are selected. Confidential

Advantages of an Industrial Placement Boosts employability- experience in specific field that will make you stand out from other candidates when it comes to applying for jobs, masters/PhD’s following graduation. Work with latest new technologies- experience with new technologies and high throughput methods such as the cytomine, AMBR 15 and Tecan. Transferrable skills- independent working, working as part of a team, time management… Networking-making contacts with professionals in industry will be useful when searching for a future job and receive unlimited career advice from someone with a lot of experience. Hands on experience in a real science environment- insight into what working in industry is actually like that you would not get from degree alone. Confidential

Questions? Confidential