Cell Biology Monoclonal Antibody Production, Cell Fusion

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1 Cell Biology Monoclonal Antibody Production, Cell Fusion This system is used for the automated cell fusion of spleen cells and SP2 cells (Myeloma Cells) for the development of cell lines which produce monoclonal antibodies. Part of the Monash system. Technical Reference Input Spleen Cells, SP2 Cells Spleen cells in 50 ml BD Falcon tubes SP2 cells in Spinner bottle continuously pumped into a 100 ml trough on the worktable Selection medium in Schott glass bottle continuously pumped into a 100 ml trough on the worktable System liquid is DMEM All other reagents in troughs on the worktable Highlights Process Red Cell Lyses, Mix Cells, Rinse, Fusion, Plating out Red cell lyses Cell count Mixing of spleen cells and SP2 cells in a specific ratio Rinse cell mixture Fusion Rinse fused cells Plating out of cells o Plating out is done in 6 well plates and 96 deep well plates Output Plates with fused Cells Fused cells in 6 well plates; 96 plates per working day Fused cells in 96 DWP; 8 DWP s per 5 hours Throughput System capacity is 24 fusions per day

2 Integration of heated Shakes for BD Falcon tubes One Shake is equipped with one electrical heated rack for 4 x 50 ml Falcon tubes. Two of these individual modules are mounted on one adapter plate to an 8 pos. carrier. The LiHa can pipette into all 8 tubes simultaneously. In the picture below there are 6 of these Shake racks visible. Worktable cutouts for the tube racks allow the Shakes being positioned below the Freedom EVO worktable. Nevertheless z-max of the tubes is equal to the surface height of the worktable. Shake 2 Shake 4 Shake 6 Shake 1 Shake 3 Shake 5 Manufactured by TIG, Tecan Schweiz AG, Seestrasse 103, 8708 Männedorf Switzerland, tig.eu@tecan.com, p 2/7

3 Lidding station for 6 well plates and 96 deep well plates The linear lidder (LiLi ) is placed over the linear transfer station of the Cytomat. The pneumatic and electronic box is placed on the right side of the LiLi. It is connected to the CAN bus of the Freedom EVO. The linear movement is driven by a motor and the lid is taken off and on by pneumatics. LiLi Transfer station Cytomat Manufactured by TIG, Tecan Schweiz AG, Seestrasse 103, 8708 Männedorf Switzerland, tig.eu@tecan.com, p 3/7

4 Manufactured by TIG, Tecan Schweiz AG, Seestrasse 103, 8708 Männedorf Switzerland, p 4/7

5 Refill Trough and peristaltic pump Ecoline V-380 Up to three 100 ml troughs can be placed onto the Refill trough carrier. For each refill trough 2 peristaltic pumps are needed: one pump pumps the liquid into the trough and one pump aspirates the excess volume from the trough. The system can also be used as recirculation trough. Refill troughs Liquid trough carrier with overflow trough Manufactured by TIG, Tecan Schweiz AG, Seestrasse 103, 8708 Männedorf Switzerland, tig.eu@tecan.com, p 5/7

6 Peristaltic Pumps Ecoline V-380 The peristaltic pumps are placed onto a shelf in the back of the Freedom EVO. Each trough has two pumps, one for input and one for output. The individual speed is set on each peristaltic pump directly. The peristaltic pumps are connected to the SMIO board of the Freedom EVO and can be switched on and off by firmware commands. Refill troughs Peristaltic pumps Manufactured by TIG, Tecan Schweiz AG, Seestrasse 103, 8708 Männedorf Switzerland, tig.eu@tecan.com, p 6/7

7 Centrifuge Buckets for 50 ml Falcon tubes Up to 4 centrifuge buckets can be loaded onto the IKA 130 Shaker. One bucket contains 6 x 50 ml Falcon tubes. The tubes can be closed with a metal lid during centrifugation. The buckets are placed into the centrifuge with a modified Giraffe extension tool which is moved by the Pick and Place (P&P) arm. Centrifuge buckets on IKA 130 Shaker Giraffe P&P arm Tube Lids Hettich centrifuge Rotanta (FE500 model) Scientific instrumentation; not for use in human, clinical or diagnostic procedure. All systems or products are tailor-made one-offs that were exclusively developed on customer demand. All displayed pictures and images originate from development-, test- or installation phases and do not claim to represent the final installation at customer site. Tecan, Freedom EVO are in major countries registered trademarks and Shake, TeLiLi and FE 500 are trademarks of Tecan Group Ltd., Männedorf, Switzerland. IKA is a registered trademark of IKA Werke GmbH Corp., Hilden, Germany. BD is a registered trademark of Becton, Dickinson and Company, Drive Franklin Lakes, USA. 2008, Tecan Trading AG, Switzerland, all rights reserved. Manufactured by TIG, Tecan Schweiz AG, Seestrasse 103, 8708 Männedorf Switzerland, tig.eu@tecan.com, p 7/7