Production Of Antibody Fragments With Plasmid-based And Genome Integrated T7 E. Coli Expression Systems: Evaluation Of Systems Performance In Microtiter Fed-batch Like Cultivations | 97609
ISSN: 2155-952X

Journal of Biotechnology & Biomaterials
Open Access

Like us on:

Our Group organises 3000+ Global Conferenceseries Events every year across USA, Europe & Asia with support from 1000 more scientific Societies and Publishes 700+ Open Access Journals which contains over 50000 eminent personalities, reputed scientists as editorial board members.

Open Access Journals gaining more Readers and Citations
700 Journals and 15,000,000 Readers Each Journal is getting 25,000+ Readers

This Readership is 10 times more when compared to other Subscription Journals (Source: Google Analytics)
Recommended Conferences

26th World Congress on Biotechnology

Prague, Czech Republic

27th World Congress on Biotechnology

Prague, Czech Republic

27th Global Congress on Biotechnology

Paris, France

29th Asia Pacific Biotechnology Congress

Dubai, UAE
Google Scholar citation report
Citations : 1820

Journal of Biotechnology & Biomaterials received 1820 citations as per Google Scholar report

Indexed In
  • Index Copernicus
  • Google Scholar
  • Sherpa Romeo
  • Open J Gate
  • Genamics JournalSeek
  • Academic Keys
  • ResearchBible
  • China National Knowledge Infrastructure (CNKI)
  • Access to Global Online Research in Agriculture (AGORA)
  • Electronic Journals Library
  • RefSeek
  • Hamdard University
  • OCLC- WorldCat
  • SWB online catalog
  • Virtual Library of Biology (vifabio)
  • Publons
  • Geneva Foundation for Medical Education and Research
  • Euro Pub
Recommended Journals
Share This Page

Production of antibody fragments with plasmid-based and genome integrated T7 E. coli expression systems: Evaluation of systems performance in microtiter fed-batch like cultivations

21st European Biotechnology Congress

Monika Cserjan, Mathias Fink, Sophie Vazulka, Johanna Jarmer and Gerald Striedner

University of Natural Resources and Life Sciences, AustriaBoehringer Ingelheim RCV GmbH & Co KG, Austria

ScientificTracks Abstracts: J Biotechnol Biomater

DOI: 10.4172/2155-952X-C5-100

Although E. coli is the most prominent bacterial production host for recombinant proteins, some proteins with high economic potential can still hardly be produced at remunerative levels. We selected four different Fabs (Fragment, antigen binding) (BIBH1, BIWA4, CIMZIA and FabX) with identical constant domains representing such challenging proteins. Fab yield can be affected by miss-folding, aggregation or unbalanced expression, translation and translocation levels of sub-units, making it still challenging to efficiently design expression systems and production processes. For translocation to the periplasm a post-translational (ompA) and a co-translational (dsbA) leader sequence were used. E. coli BL21(DE3) and E. coli HMS174(DE3) were transformed either via pET vectors or genome integration. The resulting 32 clones, were cultivated under fed-batch like conditions in the BioLector. Cell growth was not affected by leader/Fab combinations but yield of correctly folded Fab ranged from 0 to 12.5 mg/g CDM. Higher expression rates caused higher amounts of free light chain and K12 strain reached higher yields. Except of CIMZIA with the dsbA leader, genome integrated versions showed higher Fab yields, reduced levels of free light chain and basal expression than plasmid-based systems. Independent from used expression system, highest yields were obtained with CIMZIA followed by BIWA4, BIBH1 and FabX. Leader sequence cleavage-efficiency for DsbA was significantly lower than for OmpA, both showed lowest with CIMZIA. Summarizing, we showed that the selected set of host/gene dosage/leader/Fab combinations resulted in a broad range of variation in terms of Fab yields and processing and will be studied detailed in bench-scale fermentations.

Monika Cserjan has completed her PhD at the University of Natural Resources and Life Sciences, Vienna in 1998. She is Senior Scientist in the Christian Doppler Laboratory for production of next-level biopharmaceuticals in E. coli at the Department of Biotechnology (Fermentation Technology Group), Vienna and Project Leader at the Austrian Centre of Industrial Biotechnology (ACIB).

E-mail: [email protected]


Peer Reviewed Journals

Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals

Clinical & Medical Journals

Dersleri yüzünden oldukça stresli bir ruh haline sikiş hikayeleri bürünüp özel matematik dersinden önce rahatlayabilmek için amatör pornolar kendisini yatak odasına kapatan genç adam telefonundan porno resimleri açtığı porno filmini keyifle seyir ederek yatağını mobil porno okşar ruh dinlendirici olduğunu iddia ettikleri özel sex resim bir masaj salonunda çalışan genç masör hem sağlık hem de huzur sikiş için gelip masaj yaptıracak olan kadını gördüğünde porn nutku tutulur tüm gün boyu seksi lezbiyenleri sikiş dikizleyerek onları en savunmasız anlarında fotoğraflayan azılı erkek lavaboya geçerek fotoğraflara bakıp koca yarağını keyifle okşamaya başlar