1.6 Genetic Engineering of Antibodies and Derivatives from Mammalian Cells

2011 ◽  
Author(s):  
Michaela Schäffner ◽  
Brigitte Kaluza ◽  
Ulrich Η. Weidle
Keyword(s):  
Genetic Engineering ◽  
Mammalian Cells

scholarly journals Genetic engineering approach to develop next-generation reagents for endotoxin quantification

2016 ◽  
Vol 23 (2) ◽  
pp. 136-146 ◽  
Author(s):  
Hikaru Mizumura ◽  
Norihiko Ogura ◽  
Jun Aketagawa ◽  
Maki Aizawa ◽  
Yuki Kobayashi ◽  
...  
Keyword(s):  
Genetic Engineering ◽  
Mammalian Cell ◽  
Mammalian Cells ◽  
Raw Materials ◽  
Standard Curve ◽  
Future Production ◽  
Engineering Technique ◽  
Horseshoe Crabs ◽  
Factor C ◽  
Genetic Engineering Technique

The bacterial endotoxin test, which uses amebocyte lysate reagents of horseshoe crab origin, is a sensitive, reproducible and simple assay to measure endotoxin concentration. To develop sustainable raw materials for lysate reagents that do not require horseshoe crabs, three recombinant protease zymogens (factor C, derived from mammalian cells; factor B; and the proclotting enzyme derived from insect cells) were prepared using a genetic engineering technique. Recombinant cascade reagents (RCRs) were then prepared to reconstruct the reaction cascade in the amebocyte lysate reagent. The protease activity of the RCR containing recombinant factor C was much greater than that of recombinant factor C alone, indicating the efficiency of signal amplification in the cascade. Compared with the RCR containing the insect cell-derived factor C, those containing mammalian cell-derived factor C, which features different glycosylation patterns, were less susceptible to interference by the injectable drug components. The standard curve of the RCR containing mammalian cell-derived recombinant factor C had a steeper slope than the curves for those containing natural lysate reagents, suggesting a greater sensitivity to endotoxin. The present study supports the future production of recombinant reagents that do not require the use of natural resources.

Genetic Engineering in Mammalian Cells

1981 ◽  
Vol 245 (1) ◽  
pp. 106-121 ◽  
Author(s):  
W. French Anderson ◽  
Elaine G. Diacumakos
Keyword(s):  
Genetic Engineering ◽  
Mammalian Cells

Genetic engineering of mammalian cells by direct delivery of FLP recombinase protein

Methods ◽  
2011 ◽  
Vol 53 (4) ◽  
pp. 386-393 ◽  
Author(s):  
Christoph Patsch ◽  
Daniela Kesseler ◽  
Frank Edenhofer
Keyword(s):  
Genetic Engineering ◽  
Mammalian Cells ◽  
Flp Recombinase ◽  
Direct Delivery

scholarly journals DEVELOPMENT OF AUTONOMOUSLY REPLICATING VIRAL RNA TO EXPRESS THE RECOMBINANT HUMAN GRANULOCYTE COLONY-STIMULATING FACTOR

2015 ◽  
Vol 3 ◽  
pp. 398-404
Author(s):  
Makhabbat Kaliyeva ◽  
Alexander Shustov
Keyword(s):  
Cell Culture ◽  
Genetic Engineering ◽  
Viral Genome ◽  
Mammalian Cells ◽  
Viral Rna ◽  
Expression Vectors ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Mammalian Expression ◽  
Stimulating Factor

Recombinant human granulocyte colony-stimulating factor (G-CSF) produced in cultured mammalian cells undergoes proper posttranslational modifications and, thereby, possesses better pharmacological properties in comparison with the homologous protein expressed in bacteria. Biopharmaceuticals derived from cell culture tend to be expensive because of lower yields compared to bacterially expressed competitors and numerous issues with the scalability of production. Particular limitation of scalability pertains to delivery of expression vectors to the cell culture. Natural and efficient way to deliver foreign DNA or RNA to cell is a viral infection. We intended to develop the viral genome capable of G-CSF expression.Objectives: To develop autonomously replicating viral RNA capable of heterologous expression of the G-CSF in cultured mammalian cells.Methods: Genetic engineering, cell culture, and virology.Results: Viable genome of the Venezuelan equine encephalomyelitis virus (VEE) was constructed. The G-CSF gene was synthesized de novo. Gene cassette GFP-2A-G-CSF was constructed for simultaneous expression of the fluorescent marker of viral replication (GFP) and the product of interest. Recombinant viral genome VEE/GFP-2A-G-CSF was assembled; its viability was confirmed upon transfection to the BHK-21 cells.Conclusion: Genetic engineering of the cDNA copies of the genomes of RNA viruses provides excessive opportunities to develop mammalian expression systems. We achieved production of recombinant proteins in cultured mammalian cells transfected with the VEE viral RNA carrying the inserted genes of marker protein and G-CSF.

scholarly journals Use of the genetic engineering growth hormone SAISEN in children with somatotropic insufficiency: Results of clinical trials in Russia

1994 ◽  
Vol 40 (6) ◽  
pp. 30-34
Author(s):  
I. I. Dedov ◽  
V. A. Peterkova ◽  
N. P. Goncharov ◽  
T. I. Buraya ◽  
O. V. Fofanova ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Growth Rate ◽  
Clinical Trials ◽  
Side Effects ◽  
Genetic Engineering ◽  
Mammalian Cells ◽  
Treatment Duration ◽  
Laboratory Data ◽  
Efficacy And Safety ◽  
Previously Treated

The efficacy and safety of SAISEN, a recombinant human growtn hormone obtained from mammalian cells, was tested in children with hypophyseal nanism. The treatment duration was 1 year. The results indicate that SAISEN (ARES-SERONO) is a highly effective and safe preparation of growth hormone, noticeblly stimulating the growth rate both in previously untreated children with somatotropic insufficiency, and in those previously treated with STII preparations. Therapy with SAISEN was not associated with any side effects, as shown by both clinical and laboratory data.

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