miniperm Lab Scale Protein Production

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1 Cell/ 2 HTS Lab Scale Protein Production A step forward in cell culture is an economical easyto use bioreactor which simplifies high density cultivation of hybridoma and other cell types. The main focus is laid on cost efficient production of antibodies and other desired extracellular products. is suitable for cultivation of a variety of cell types. High densities of both suspension cells and anchorage dependent cells can be cultivated in the. is an effective alternative to static culture, roller bottles and fermentor systems because high concentrations of cell products can be obtained even in small volumes. is appropriate for use in a wide spectrum of research and laboratory applications. In addition to cultivation of hybridoma for antibody production and transfected cells for protein production, is also applicable for virus production and general proliferation of biomass. Depending on the cells used, cell densities up to 0 8 cells/ml and product concentrations of several mg/ml can be achieved. Special adaptation of culture techniques to is not necessary. is suitable for serumcontaining and serumfree culture. 5 Tubes/Multi Construction and Principle construction is a twocompartment bioreactor. The production module (with culture volumes of 35 ml) is separated from the (400 ml working volume) by a dialysis membrane (Fig. ). Basic principle When the production and s are assembled, the semipermeable dialysis membrane forms the dividing wall between the two modules. Neither the cells, nor high molecular weight products released by the cells are able to pass through the membrane (Fig. 2). However, nutrients and dissolved gases can diffuse from the into the. The system The is available in two versions: The assembled bioreactor, sterile: The production and s are delivered assembled and sterilized. This represents a singleuse product. The reusable : The can be autoclaved and reused. The is sterile and can be used only once. glucose lactate Figure 0 Biochips/ Cryo CO 2 cell O 2 Figure 2: Principle of NH 4 dialysis membrane cell product silicone membrane Cultivation In the the nutrient medium as well as the cell suspension must be rotated constantly, preferably on the Universal Turning Device (Fig. 3). Then cells are optimally supplied and their metabolic waste products are removed. The Universal Turning Device provided for use with enables up to four bioreactors to be simultaneously rotated. In order to achieve optimal gasexchange, the bioreactor is rotated on the Universal Turning Device inside a CO 2 incubator with a relative humidity of at least 70 %. 2 Lids/Sealers/ 3 Reaction Tubes/ 4 Accessories At the same time, low molecular weight metabolites secreted by the cells can leave the via the dialysis membrane where they are diluted in the medium of the nutrient module. The offers a large ph buffering capacity. The outer side of the is sealed with a thin, gaspermeable silicone membrane for an optimal gas exchange (O 2 and CO 2 ). The screw cap of the with an integrated membrane enables equilibration of pressure between the bioreactor and the outer environment Figure 3: Universal Turning Device

2 Cell/ 2 HTS New Selecting the Production Module NonPyrogenic Advantages High cell densities High product concentrations Continuous harvest Long term cultivation Ease of use Wide application spectrum Production of proteins/ monoclonal antibodies Cultivation of Hybridoma Mammalian cells Production of biomass Transfected cells Production of viruses Plant cells Microorganisms 5 Tubes/Multi classic The classic has a culture volume of 35 ml which is ideal for use in research and labscale applications. Suspended cells or adherent cells on microcarriers can be cultivated in the classic. Whilst the is optimized for the cultivation of hybridoma cells, many other cell types (e.g. CHO, BHK, SF, tumour cells etc.) can be successfully cultured at high densities as well. SM Stationary matrix for adherent cells The SM is ideal for the cultivation of anchorage dependent cells (e.g. HEK or COS cells). These modules contain two polyester matrices upon which many cell types attach and divide. The SM s have a growth surface area of 240 cm 2. Cell products are normally released into the production module medium. In applications for the proliferation of biomass, the stationary matrix enables a simple biomass harvesting at the end of a run as the cells are retained upon the matrix ( Fig. ). Figure noncytotoxic classic Kit classic Test Kit Production Module classic NMWCO* of Membrane [kda] bioreactors bioreactors Biochips/ Startup Support Kit Cryo NMWCO* of Membrane [kda] SM 205 Kit 2.5 SM 205 Test Kit 2.5 Production Module SM Lids/Sealers/ * nominal molecular weight cutoffs 2 bioreactors 4 bioreactors Startup Support Kit 2 3 Reaction Tubes/ Accessories

3 Accessories Cell/ 2 HTS The following accessories are offered for convenient handling during cultivation: Tubes/Multi singleuse 50 ml Luer lock and 2 ml Luer syringes (Cat. No , ) For simple and safe cell suspension inoculation, sampling and harvesting. filling tube 5'' (Cat.No ) The sterile filling tube offers a simple solution to cell suspension transfer (e.g. from centrifuge tubes to singleuse syringe). The filling tube is slightly flexible, connects directly to a 50 ml Luer syringe and has a sufficiently large inner diameter to minimize shearstress induced cell damage. Luer syringe needles (25Gx 5/8'') (Cat.No ) For sampling and supplementing the medium in the production module via septum ports. Septum port (Cat.No ) To simplify sampling and supplementing procedures and to further reduce contamination risks, a replacement of two of the three standard screw caps on the with sterile septum ports is possible. The rubber septum ports offer the possibility for addition of fluids to or removal of samples from the using sterile singleuse syringe needles of small diameter. There is no need to open the module. The septum ports should be replaced after they have been pierced up to 0 times. stand (Cat.No ) For mounting the bioreactor during inoculation, sampling and harvest. cellprotect (Cat.No ) cellprotect increases the viscosity of the culture medium and thereby protects the cells against shear stress. cellprotect is added to the cell suspension in the at concentrations of 0.05 to 0. % w/v. antifoam a (Cat.No ) When serumcontaining medium is used, significant quantities of foam may develop in both modules of the. By adding 0.5 to ml antifoam a to the medium, this foam buildup is reduced. Startup Support Kit Accessories to inoculate, sample and harvest. Consists of: Filling tube 5", Luer, sterile (8x) Disposable hypodermic syringe 50 ml Luer, sterile (8x) Disposable hypodermic syringe 2 ml Luer, sterile (20x) Luer syringe needles (25G x 5/8"), sterile (20x) Septum ports, sterile (6x) stand (x) antifoam a, 5 ml, sterile (x) cellprotect, ml, sterile (x) 3 Reaction Tubes/ 2 Lids/Sealers/ 0 Biochips/ 4 Accessories Cryo 34

4 Cell/ Nutrient module HTS Autoclavable Stand for 4 Septum port 00 Filling tube 5'' (27 mm) Luer 00 5 Tubes/Multi , singleuse , singleuse Luer syringe needles (25 G x 5/8'') syringe 2 ml Luer syringe 50 ml Luer Lock Startup Support Kit antifoam a cellprotect Volume [ml] * *except stand 3 Reaction Tubes/ 2 Lids/Sealers/ Cryo 0 Biochips/ Accessories

5 Cell/ 2 HTS Turning / Rocking Device Turning /Rocking Device 5 Tubes/Multi New Universal Turning Device A turning device for universal application In contrast to standard turning devices, the speed of this Universal Turning Device is digitally regulated across a rotational speed range of 0. to 40 rpm in steps of 0. rpm. It is therefore suitable for most laboratory applications. The Universal Turning Device can be used with almost all laboratory incubators. The control unit remains outside the incubator and is connected to the Universal Turning Device by a flat cable that passes through the door seal. Flexibility is guaranteed because the distance between the rollers is variable. It is therefore possible to simultaneously rotate different types of devices, e.g. roller bottles on one side and centrifuge tubes on the other. Special applications The Universal Turning Device is ideal for high density cell culture in the bioreactor because the rotational speed can be accurately regulated over a wide range. Optimal conditions for supplying the cells and removing their metabolic waste products and optimal gas exchange can be achieved. Universal Turning Device Advantages Flexible application Space saving Incremental rotational speed control Simple installation bioreactors Roller bottles Centrifuge tubes Reagent vessels The Rocking Device Rocking Device Advantages Simple setup Different angles of inclination Cell culture vessels (e.g. quadriperm, TFlasks, lumox TM dish, petri dishes) Protein/DNAstaining Rocking Device Various static cell culture vessels can be mounted on the Rocking Device which provides a work surface of 30 cm x 23 cm. Via the adapter, two different angles of inclination (3 and 5 ) can be set. The rocking frequency can be regulated between 0. and 40 Hz Biochips/ Cryo External dimensions [cm] Universal Turning Device 45x0x25 Rocking Device 45x0x25 Weight [kg] Lids/Sealers/ Rated voltage [V] Rated fequency [Hz] Turning speed [rpm] 240/ / / /60 3 Reaction Tubes/ 4 Accessories 36

6 Cell/ Accessories for the Universal Turning Device 2 HTS 5 Tubes/Multi 0 Biochips/ Cryo 3 Reaction Tubes/ 2 Lids/Sealers/ New Enhanced flexibility Special adapters are available for the use of centrifuge tubes and reagent vessels in conjunction with the Universal Turning Device. Three versions of adapter are available: one for up to five 50 ml centrifuge tubes, one for twelve 5 ml tubes and one with a capacity to rotate 48.5 ml/2 ml tubes Adapter for centrifuge tubes, 2 x 5 ml Adapter for centrifuge tubes, 5 x 50 ml Adapter for reagent vessels, 48 x.5/2.0 ml Spare parts for Universal Turning Device Motor Drive roller Slave roller Spacer Tray Control unit 2 240/20 V / 50/60 Hz Accessories