Navigation Links
Gram Quantities of MAb Produced with Simple Bioreactor in Serum-Free Perfusion Culture Replacing Ascitic Fluid Production

By J. Doyle, L. Johnstone and L. Cottis
AGEN Biomedical Ltd. Queensland, Australia
http://www.agen.com.au


The CelliGen Plus immobilized-bed perfusion was investigated as a means for producing large quantities of MAb from a hybridoma, as an alternative to in vivo production. This system best represented a production facility that was easy to operate, flexible in its use, small enough to enable autoclaving and yet large enough to produce the required quantities of MAb. Three cell culture trials were performed using serum-free medium. Each run proceeded in the same manner, with MAb yields of 39.3, 33.8 and 30.3 grams. The perfusion system enabled us to scale down the size of the reactor needed, as the yields were significantly higher than that obtainable in batch fermentation which would have necessitated a bioreactor of several hundred liters.


Materials and Methods
All runs were performed in a 7.5 Liter NBS CelliGen Plus bioreactor with the basket impeller (New Brunswick Scientific, Edison, NJ). Working volume was 5 L and the bed volume was 2.6 L. Cells were inoculated at a cell density of 2 x 105 cells/mL at > 80% viability. Temperature was maintained at 37C, pH 7.20 and top dissolved oxygen (DO) at 50% of air saturation. The pH was controlled via the interactive pH-DO controller with CO2 at the initial stage and then with an 8% NaHCO3 solution when perfusion reached 2 volumes/day. Agitation rate was initiated at 80 rpm and increased throughout the run to maintain the bottom DO concentration at > 35% air saturation. The maximal rate did not exceed 150 rpm. Gas supply was via the ring sparger at a rate not exceeding 0.13 LPM and this was only increased when the top DO setpoint could not be maintained.

Gibcos Hybridoma Serum-Free Medium was used, supplemented with between 2 - 4 mM L-glutamine and 100 U/mL penicillin and 100 Ug/mL streptomycin. A separate 5% glucose solution was fed to the cultures when the perfusion rate reached 2 volumes/day to maintain the residual glucose concentration at > 1 g/L. The hybridoma cell line used produces an IgG1 MAb. Perfusion at 1 volume/ day (based on the bed volume), was initiated 24 hours after the commencement of the run, with increases to 2 then 3 volumes/day also 24 hours apart. The perfusion rate was maintained at 3 volumes/day for the duration.


Results and Discussion
The MAb produced in these cultivations has been traditionally produced from ascitic fluid with an average yield after purification of 5 mg/mouse. Yields produced using a contract service in 10 & 250 L batch reactors produced ~35 mg/L before purification. Using the CelliGen Plus perfusion system, yields of between 4 - 5 times that of the batch fermentation were obtained, based on purified material from the perfusion system but unpurified material from the batch system ( see table ).

All AGEN staff found the CelliGen Plus bioreactor very user friendly, easily simplifying an otherwise complicated and laborious procedure. The bioreactors display and associated BioCommand software provided fermentation information at the touch of a fingertip and trend graphing of process controls could also be viewed via BioCommand software. These features made the data handling, interpreting and report writing much simpler, something much appreciated by all staff. For CelliGen Plus literature contact your local NBS sales representative or write us at bioinfo@nbsc.com.



'"/>

Source:


Page: All 1 2

Related biology technology :

1. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
2. A Simple and Rapid Technique to Purify High-Quality Midiprep DNA
3. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
4. Simple, Sensitive Isotyping of Mouse Monoclonal Antibodies
5. Simple Isolation of RNA from Tissue and Cultured Cells
6. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
7. Simple and Rapid Isolation of Plant RNA Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit
8. The Fastest, Simplest, and Most Effective Way to Remove DNA Contamination
9. Simple Approaches for Optimization of RT-PCR
10. Detection of Morphine and Morphine-Glucuronide in Saliva or Urine with a Rapid and Simple Method Without Derivatization on the Varian 500-MS
11. CHO Cell Culture Application Using a BioFlo 110 Benchtop Bioreactor
Post Your Comments:
(Date:8/31/2015)... MARLBOROUGH, Mass. , Aug. 31, 2015 /PRNewswire/ ... biotechnology company focused on discovering and developing innovative ... ophthalmology, today announced that the 30-day review period ... Drug (IND) for ocular indications with the U.S. ... the IND is now active. ...
(Date:8/31/2015)... HAYWARD, Calif. and SHANGHAI ... Inc., a privately-held biopharmaceutical company developing next-generation antibiotics, ... Phase 2 clinical study for its lead drug ... designed to treat drug-resistant bacteria such as MRSA ... safer and better tolerated therapeutic option than currently ...
(Date:8/31/2015)... 31, 2015 LabStyle Innovations Corp. (OTCQB: ... today announced it recently closed a round of ... used to support the global rollout and initial ... cloud-based, diabetes management solution that includes novel software ... glucose monitoring device, and for working capital purposes. ...
(Date:8/31/2015)... Grove, IL (PRWEB) , ... August 31, 2015 ... ... impurities (PGIs), as a high priority in the drug development approval process. Thus, ... , This year, Regis Technologies, Inc., a Chicago-based CMO, has been hosting ...
Breaking Biology Technology:RXi Pharmaceuticals Announces Advancements in its Ophthalmology Clinical Program with RXI-109 2RXi Pharmaceuticals Announces Advancements in its Ophthalmology Clinical Program with RXI-109 3RXi Pharmaceuticals Announces Advancements in its Ophthalmology Clinical Program with RXI-109 4MicuRX Reports Positive Top-Line Results In Phase 2 Clinical Trial For Novel Antibiotic MRX-I In Complicated Skin And Soft Tissue Infections 2LabStyle Innovations Closes $2.5 Million Funding to Support Global Rollout of Dario Diabetes Management Solution 2LabStyle Innovations Closes $2.5 Million Funding to Support Global Rollout of Dario Diabetes Management Solution 3LabStyle Innovations Closes $2.5 Million Funding to Support Global Rollout of Dario Diabetes Management Solution 4Regis Shares Expert Advice on Impurities in Drug Development 2Regis Shares Expert Advice on Impurities in Drug Development 3
... Thailand, Sept. 8 Don Margolis, Chairman of,The ... ) based in,Bangkok, Thailand told The Wall Street ... that future profits inside the stem cell,research sector ... next five years, even for,companies specializing in Repair ...
... PRINCETON, New Jersey and SAN DIEGO,California, September 8 , ... for Continuous Process,Chemistry Technologies and for Further Global Expansion, ... investment of $3.3M,by a group of private and financial ... Graphisoft and Dave Tapolczay, CEO of Medical,Research Council Technology ...
... Sept. 8 /PRNewswire-FirstCall/,-- Concuity, a healthcare division ... a leading,provider of contract management and revenue ... today announced that Valley Baptist Health,System has ... more,accurately calculate payments from managed care and ...
Cached Biology Technology:Stem Cell Profits Five Years Away, Says Repair Stem Cell Institute Chairman to The Wall Street Transcript 2World Leader in Bench-Top Flow Chemistry Instruments ThalesNano, Inc. Closes Funding Round 2Valley Baptist Health System Selects Concuity's ClearContracts to More Accurately Calculate Payments from Managed Care and Government Payers 2Valley Baptist Health System Selects Concuity's ClearContracts to More Accurately Calculate Payments from Managed Care and Government Payers 3Valley Baptist Health System Selects Concuity's ClearContracts to More Accurately Calculate Payments from Managed Care and Government Payers 4
(Date:8/5/2015)... , August 5, 2015 ... titled "Facial Recognition Market - Global Industry Analysis, Size, ... global market for facial recognition is forecast to reach ... by increased demand for surveillance systems by civil and ... number of crimes and terrorist activities across the globe ...
(Date:8/3/2015)... 2015 Synaptics, Inc. (Nasdaq: SYNA ), ... that members of the executive management team will present ... Technology Leadership Forum on Tuesday, August 11, 2015 at ... at the Sonnenalp Resort in Vail, CO. ... forward-looking information. An audio webcast of the event will ...
(Date:7/31/2015)... La 10e Conférence internationale sur la ... le BGI du 22 au 25 octobre 2015 ... La conférence célèbre son 10e anniversaire ... est devenue l,une des réunions annuelles mondiales les ... c,est aussi la plus dynamique, la plus enthousiaste ...
Breaking Biology News(10 mins):Facial Recognition Market Expected to Reach US$ 2.67 Bn by 2022, at a CAGR of 9.5% From 2015 - 2022: Transparency Market Research 2Facial Recognition Market Expected to Reach US$ 2.67 Bn by 2022, at a CAGR of 9.5% From 2015 - 2022: Transparency Market Research 3Facial Recognition Market Expected to Reach US$ 2.67 Bn by 2022, at a CAGR of 9.5% From 2015 - 2022: Transparency Market Research 4Facial Recognition Market Expected to Reach US$ 2.67 Bn by 2022, at a CAGR of 9.5% From 2015 - 2022: Transparency Market Research 5Facial Recognition Market Expected to Reach US$ 2.67 Bn by 2022, at a CAGR of 9.5% From 2015 - 2022: Transparency Market Research 6La 10e International Conference on Genomics (ICG-10) doit s'ouvrir en octobre 2La 10e International Conference on Genomics (ICG-10) doit s'ouvrir en octobre 3
... a grant of $8,950,590 provided through the American Recovery and ... Diego looking for the biological bases of differences in human ... collect a wealth of data about brain development in children. ... Institute on Drug Abuse (NIDA), a part of the National ...
... authorized more than $29 million for the University of ... a former gold mine in South Dakota into the ... by the National Science Board, is for a preliminary ... (DUSEL), a facility at the former Homestake gold mine ...
... including a University of Florida paleontologist has used a ... provide the first reliable evidence of how Neotropical rainforests ... Smithsonian Institution and UF, among others, found that many ... rainforests including legumes, palms, avocado and banana ...
Cached Biology News:Stimulus grant of nearly $9 million to UC San Diego funds big study of young brains 2Stimulus grant of nearly $9 million to UC San Diego funds big study of young brains 3NSF authorizes $29 million for preliminary design of world's deepest underground laboratory 2NSF authorizes $29 million for preliminary design of world's deepest underground laboratory 3Plant fossils give first real picture of earliest Neotropical rainforests 2Plant fossils give first real picture of earliest Neotropical rainforests 3
... Timp-4 has been reported in heart ... fibroblasts in cell culture conditions. ... TPA stimulates production of TIMP-4 in ... protein levels produced often require concentration ...
Goat polyclonal to HSPC150 ( Abpromise for all tested applications). entrezGeneID: 29089...
Anti-MMP-13 Collagenase-3, amino terminal end of active enzyme; rabbit host...
...
Biology Products: