Abstract:
: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.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Tutykhina IL,Bezborodova OA,Shmarov MM,Logunov DY,Neugodova GL,Nemtsova ER,Naroditsky BS,Yakubovskaya RI,Gintsburg ALdoi
10.1016/j.pep.2009.01.003subject
Has Abstractpub_date
2009-05-01 00:00:00pages
100-7issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(09)00002-3journal_volume
65pub_type
杂志文章abstract::Bikunin is a proteoglycan exhibiting broad-spectrum inhibitory activity against serine proteases and could potentially suppress tumor cell invasion and metastasis. Here, we have successfully expressed recombinant human bikunin (rh-bikunin) in the methylotrophic yeast Pichia pastoris and established the purification pr...
journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
pub_type: 杂志文章
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abstract::Recombinant human interleukin-5 (rhIL-5) was expressed in baculovirus-infected insect cells and purified to homogeneity from the culture medium in a single chromatographic step. Beginning with a cDNA encoding the full-length precursor form of human IL-5, including the authentic secretory leader sequence, recombinant b...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.1009
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journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1995.1077
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journal_title:Protein expression and purification
pub_type: 杂志文章
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0103
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journal_title:Protein expression and purification
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abstract::The methylotrophic yeasts Pichia pastoris and Pichia angusta (Hansenula polymorpha) were used for the comparative heterologous production of two model mammalian proteins of pharmaceutical interest, the NK1-fragment (22 kDa) of human hepatocyte growth factor and the extracellular domain (28 kDa) of mouse tissue factor ...
journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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journal_title:Protein expression and purification
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abstract:UNLABELLED:Recombinant protein production is a significant biotechnological process as it allows researchers to produce a specific protein in desired quantities. Escherichia coli (E. coli) is the most popular heterologous expression host for the production of recombinant proteins due to its advantages such as low cost,...
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