Biomaterial Database

Production of recombinant human lactoferrin in the allantoic fluid of embryonated chicken eggs and its characteristics. 2009

Irina L Tutykhina, and Olga A Bezborodova, and Maxim M Shmarov, and Denis Y Logunov, and Galina L Neugodova, and Elena R Nemtsova, and Boris S Naroditsky, and Raisa I Yakubovskaya, and Alexander L Gintsburg

Laboratory of Molecular Biotechnology, Gamaleya Research Institute of Epidemiology and Microbiology, 18, Gamaleya Street, Moscow 123098, Russia.

The human iron-binding protein lactoferrin (hLf) has been implicated in a number of important physiological pathways, including those regulating immune function and tumor growth. In an effort to develop an efficient system for production of recombinant hLf (rhLf) that is structurally and functionally equivalent to the natural protein, we generated a recombinant CELO (chicken embryo lethal orphan) avian adenovirus containing an expression cassette for hLf. Embryonated chicken eggs were infected with the generated CELO-Lf virus. rhLf expression was measured in the allantoic fluid of infected eggs by ELISA three days later. The level of recombinant protein was about 0.8mg per embryo. rhLf was efficiently purified (up to 85% yield) from the allantoic fluid of infected eggs using affinity chromatography. rhLf produced in the allantoic fluid was characterized in comparison with natural hLf (nhLf) purified from human breast milk. SDS-PAGE, Western blotting and glycosylation analyzes showed that the recombinant protein had similar physical characteristics to nhLf. In addition, we demonstrated that the antioxidative and antimicrobial activity of rhLf produced in this system is equivalent to that of nhLf. Taken together, these results illustrate the utility of the described "recombinant CELO adenovirus-chicken embryo" system for production of functionally active rhLf. Efficient production of rhLf with accurate structure and function is an important step in furthering investigation of Lf as a potential human drug.

UI	MeSH Term	Description	Entries
D007781	Lactoferrin	An iron-binding protein that was originally characterized as a milk protein. It is widely distributed in secretory fluids and is found in the neutrophilic granules of LEUKOCYTES. The N-terminal part of lactoferrin possesses a serine protease which functions to inactivate the TYPE III SECRETION SYSTEM used by bacteria to export virulence proteins for host cell invasion.	Lactotransferrin
D011994	Recombinant Proteins	Proteins prepared by recombinant DNA technology.	Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002352	Carrier Proteins	Proteins that bind or transport specific substances in the blood, within the cell, or across cell membranes.	Binding Proteins,Carrier Protein,Transport Protein,Transport Proteins,Binding Protein,Protein, Carrier,Proteins, Carrier
D002642	Chick Embryo	The developmental entity of a fertilized chicken egg (ZYGOTE). The developmental process begins about 24 h before the egg is laid at the BLASTODISC, a small whitish spot on the surface of the EGG YOLK. After 21 days of incubation, the embryo is fully developed before hatching.	Embryo, Chick,Chick Embryos,Embryos, Chick
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000259	Aviadenovirus	A genus of ADENOVIRIDAE that infects birds. The type species is FOWL ADENOVIRUS A.	Adenoviruses, Avian,Avian Adenoviruses,Adenovirus, Avian,Aviadenoviruses,Avian Adenovirus
D000482	Allantois	An extra-embryonic membranous sac derived from the YOLK SAC of REPTILES; BIRDS; and MAMMALS. It lies between two other extra-embryonic membranes, the AMNION and the CHORION. The allantois serves to store urinary wastes and mediate exchange of gas and nutrients for the developing embryo.	Allantoic Membrane,Membrane, Allantoic
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D015053	Zygote	The fertilized OVUM resulting from the fusion of a male and a female gamete.	Fertilized Ovum,Ovum, Fertilized,Fertilized Egg,Egg, Fertilized,Eggs, Fertilized,Fertilized Eggs,Zygotes
D015870	Gene Expression	The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.	Expression, Gene,Expressions, Gene,Gene Expressions