Abstract
The tubular gland of the chicken oviduct is an attractive system for protein expression as large quantities of proteins are deposited in the egg, the production of eggs is easily scalable and good manufacturing practices for therapeutics from eggs have been established. Here we examined the ability of upstream and downstream DNA sequences of ovalbumin, a protein produced exclusively in very high quantities in chicken egg white, to drive tissue-specific expression of human mAb in chicken eggs. To accommodate these large regulatory regions, we established and transfected lines of chicken embryonic stem (cES) cells and formed chimeras that express mAb from cES cell-derived tubular gland cells. Eggs from high-grade chimeras contained up to 3 mg of mAb that possesses enhanced antibody-dependent cellular cytotoxicity (ADCC), nonantigenic glycosylation, acceptable half-life, excellent antigen recognition and good rates of internalization.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Antibodies, Monoclonal / chemistry*
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Blotting, Southern
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Blotting, Western
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CHO Cells
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Calorimetry, Differential Scanning
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Carbohydrates / chemistry
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Chickens
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Cricetinae
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DNA / metabolism
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Egg White
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Embryo, Mammalian / cytology
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Embryo, Nonmammalian
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Enzyme-Linked Immunosorbent Assay
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Female
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Genetic Vectors
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Genome
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Glycosylation
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Humans
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Immunoglobulin G
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Immunohistochemistry
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Isoelectric Focusing
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Mice
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Mice, Inbred BALB C
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Models, Genetic
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Monosaccharides / chemistry
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Oligosaccharides / chemistry
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Ovalbumin / genetics
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Ovalbumin / metabolism
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Polymerase Chain Reaction
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Recombinant Fusion Proteins / chemistry
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Reverse Transcriptase Polymerase Chain Reaction
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Spectrometry, Mass, Electrospray Ionization
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Stem Cells / cytology
Substances
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Antibodies, Monoclonal
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Carbohydrates
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Immunoglobulin G
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Monosaccharides
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Oligosaccharides
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Recombinant Fusion Proteins
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Ovalbumin
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DNA