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Newsletters

Periodic bite-sized updates and information on hollow fiber

Newsletters

Latest newsletter

04/29/2021 – FiberCell Systems Expressions – April 2021 Newsletter

Past newsletters

02/24/2021 – FiberCell Systems Expressions – February 2021 Newsletter
08/26/2020 – FiberCell Systems Expressions – August 2020 Newsletter
08/12/2020 – First-time User of 3-D Capillary Bioreactor Produces Nearly 2 Grams of Antibody
07/09/2020 – FiberCell Systems Expressions – July 2020 Newsletter
06/09/2020 – MINI-WEBINAR:Beyond 3-D: Recapitulating Nature with a Hollow Fiber Bioreactor
06/02/2020 – FiberCell Systems Expressions – June 2020 Newsletter
05/21/2020 – MINI-WEBINAR:The Hollow Fiber Infection Model and Antibiotic Pk/Pd
05/21/2020 – MINI-WEBINAR:Clinical Scale Production of Extracellular Vesicles in a Hollow Fiber Bioreactor System
05/21/2020 – MINI-WEBINAR: Production of Difficult-to-Express Proteins And Monoclonal Antibodies in Hollow Fiber Bioreactors
03/11/2020 – Hollow Fiber Bioreactors and COVID-19
11/26/2019 – Produce GRAM QUANTITIES of Recombinant Proteins Expressed in Mammalian Cells in a Hollow Fiber Bioreactor
02/12/2019 – Winter 2019 Expressions Newsletter from FiberCell Systems
09/26/2018 – FiberCell Expressions Newsletter September 2018
10/19/2017 – FiberCell Expressions Newsletter October 2017
07/25/2017 – FiberCell Expressions Newsletter July 2017
07/13/2017 – FiberCell Expressions Newsletter July 2017
01/31/2017 – FiberCell Expressions Newsletter January 2017
10/25/2016 – FiberCell Expressions Newsletter October 2016

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Simply perform your harvest as usual. One of the issues that many researchers have when using our system is that they are afraid to harvest out too many cells. The cartridge can be overgrown if you don’t remove cells. After centrifugation your cell pellet should be between 1-3 mL or more (with the C5011 as much as 5 mL). Simply decant off the supernatant and re-suspend the cell pellet in 2 pellet volumes of fresh medium that has been diluted 10% with sterile distilled water. Leave it overnight at room temperature. The cells will consume the medium and also the hypotonicity will slightly squeeze the cells causing them to release any intracellularly sequestered antibody. This will get you an extra few milligrams every time you harvest.

Too few cells in relation to volume of media can result in lag phase. In the hollow fiber cartridge lag phase can be defined by the cells not consuming glucose but still show significant viability via the trypan exclusion assay. Always seed the cartridge with the recommended cell density. If this is not possible, adjust the volume of media down to keep the cells in culture from being too dilute.If the cells do go into lag phase you can either:

reduce the volume in the reservoir bottle back down to 125 mL
inoculate more cells
wait a few days – sometimes the trick with hollow fiber cell culture is to know when to NOT do something.

Hybridoma cells growing at high density in the C2011 cartridge using CDM-HD medium

To keep contamination at a minimum, clean the inside of the hood weekly, remembering to remove the working surface and clean underneath. Be sure the hood is validated on a regular basis. Never pour media, always pipette. Always wear a lab coat. Wait 60 seconds for the air currents to stabilize inside the incubator. Pull hair back and be sure to never have bare flesh inside the hood. Use plenty of alcohol and wipes but remember, it is not the application of alcohol that sterilizes, it is the evaporation. Good sterile technique will allow production from the cartridge for many months or more. We have produced a monoclonal antibody for over one year of continuous production.

New cell lines should always be quarantined until a mycoplasma screen has shown negative. The presence of mycoplasma contamination can be hard to detect without specific assays. The culture will just appear to be non-productive.

Plama-Pie1

Various components of serum. The major component, BSA binds many substances non-specifically and can be an overwhelming protein burden in some purification schemes, immunoglobulins can interfere with specific antibody purifications and the “other” can have many undefined effects. None of these characteristics will be reproducible from lot to lot.