An integrated vector system for the eukaryotic expression of antibodies or their fragments after selection from phage display libraries.
Phage display is now an established method to select antibody fragments specific for a wide range of diverse antigens. In particular, isolation of human monoclonal antibodies has become a reality and for most purposes bacterial expression of the selected recombinant antibody fragments is sufficient. However, there are some cases where the expression of complete human immunoglobulin in mammalian cells is, if not essential, at least desirable. For this reason we have designed and constructed a set of mammalian expression vectors which permit facile and rapid cloning of antibody genes for both transient and stable expression in mammalian cells. Immunoglobulin genes may be cloned into these expression vectors as V regions or as Fabs for expression as either complete antibodies or as Fab fragments, using restriction sites which are rare in human V genes. All the important elements in the vectors--promoter, leader sequence, constant domains and selectable markers--are flanked by unique restriction sites, allowing simple substitution of elements. The vectors have been evaluated using the variable regions from the neutralizing anti-nerve growth factor (NGF) antibody, alphaD11, and the V regions from 2E10, a scFv selected from a scFv phagemid library.
Persic
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Societa Italiana per la Ricerca Scientifica, Roma, Italy.
Roberts
A
A
Wilton
J
J
Cattaneo
A
A
Bradbury
A
A
Hoogenboom
H R
HR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D001435:Bacteriophages; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D017931:DNA Primers; D005803:Genes, Immunoglobulin; D005822:Genetic Vectors; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology; D009414:Nerve Growth Factors / Q000276:immunology; D019151:Peptide Library; D010957:Plasmids; D011994:Recombinant Proteins / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D009414:Nerve Growth Factors; D019151:Peptide Library; D011994:Recombinant Proteins