scholarly journals piggyBac Transposition and the Expression of Human Cystatin C in Transgenic Chickens

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1554
Author(s):  
Seo Woo Kim ◽  
Jeong Hyo Lee ◽  
Ji Seon Han ◽  
Seung Pyo Shin ◽  
Tae Sub Park

A bioreactor can be used for mass production of therapeutic proteins and other bioactive substances. Although various methods have been developed using microorganisms and animal cells, advanced strategies are needed for the efficient production of biofunctional proteins. In microorganisms, post-translational glycosylation and modification are not performed properly, while animal cell systems require more time and expense. To overcome these problems, new methods using products from transgenic animals have been considered, such as genetically modified cow’s milk and hen’s eggs. In this study, based on a non-viral piggyBac transposition system, we generated transgenic bioreactor chickens that produced human cystatin C (hCST3). There were no differences in the phenotype or histochemical structure of the wild-type and hCST3-expressing transgenic chickens. Subsequently, we analyzed the hCST3 expression in transgenic chickens, mainly in muscle and egg white, which could be major deposition warehouses for hCST3 protein. In both muscle and egg white, we detected high hCST3 expression by ELISA and Western blotting. hCST3 proteins were efficiently purified from muscle and egg white of transgenic chickens using a His-tag purification system. These data show that transgenic chickens can be efficiently used as a bioreactor for the mass production of bioactive materials.

FEBS Letters ◽  
1988 ◽  
Vol 236 (1) ◽  
pp. 14-18 ◽  
Author(s):  
Magnus Abrahamson ◽  
Henrik Dalbøge ◽  
Isleifur Olafsson ◽  
Søren Carlsen ◽  
Anders Grubb

Genes ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 38
Author(s):  
Takehiro Mukae ◽  
Sho Okumura ◽  
Takuma Watanobe ◽  
Kyoko Yoshii ◽  
Takahiro Tagami ◽  
...  

Increased commercial demand for monoclonal antibodies (mAbs) has resulted in the urgent need to establish efficient production systems. We previously developed a transgenic chicken bioreactor system that effectively produced human cytokines in egg whites using genome-edited transgenic chickens. Here, we describe the application of this system to mAb production. The genes encoding the heavy and light chains of humanized anti-HER2 mAb, linked by a 2A peptide sequence, were integrated into the chicken ovalbumin gene locus using a CRISPR/Cas9 protocol. The knock-in hens produced a fully assembled humanized mAb in their eggs. The mAb expression level in the egg white was 1.4–1.9 mg/mL, as determined by ELISA. Furthermore, the antigen binding affinity of the anti-HER2 mAb obtained was estimated to be equal to that of the therapeutic anti-HER2 mAb (trastuzumab). In addition, antigen-specific binding by the egg white mAb was demonstrated by immunofluorescence against HER2-positive and -negative cells. These results indicate that the chicken bioreactor system can efficiently produce mAbs with antigen binding capacity and can serve as an alternative production system for commercial mAbs.


1997 ◽  
Vol 11 (1) ◽  
pp. 111-118 ◽  
Author(s):  
Paul J. Berti ◽  
Irena Ekiel ◽  
Peter Lindahl ◽  
Magnus Abrahamson ◽  
Andrew C. Storer

Amino Acids ◽  
2016 ◽  
Vol 48 (12) ◽  
pp. 2809-2820 ◽  
Author(s):  
Martyna Prądzińska ◽  
Izabela Behrendt ◽  
Juan Astorga-Wells ◽  
Aleksandr Manoilov ◽  
Roman A. Zubarev ◽  
...  

1993 ◽  
Vol 22 (4) ◽  
pp. 697-701 ◽  
Author(s):  
Juan Antonio Garc�a ◽  
Mar�a Teresa Cervera ◽  
Jos� Luis Riechmann ◽  
Carlos L�pez-Ot�n

2018 ◽  
Vol 123 (17) ◽  
pp. 174701 ◽  
Author(s):  
Magdalena Chrabąszczewska ◽  
Martyna Maszota-Zieleniak ◽  
Zuzanna Pietralik ◽  
Michał Taube ◽  
Sylwia Rodziewicz-Motowidło ◽  
...  

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