An integrated vector system for the eukaryotic expression of antibodies or their fragments after selection from phage display libraries
| Autor: | Lidija Persic, Hennie R. Hoogenboom, Antonino Cattaneo, Jane Wilton, Andrew Bradbury, Andy Roberts |
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| Přispěvatelé: | Persic, L, Roberts, A, Wilton, J, Cattaneo, Antonino, Bradbury, A, Hoogenboom, H. R. |
| Rok vydání: | 1997 |
| Předmět: |
Phage display
medicine.drug_class Phagemid Genetic Vectors Immunoglobulin Variable Region Cloning vector CHO Cells Computational biology Biology Transfection Monoclonal antibody Immunoglobulin Fab Fragments Peptide Library Cricetinae Genetics medicine Animals Humans Bacteriophages Nerve Growth Factors Cloning Molecular Immunoglobulin Fragments Gene Selectable marker DNA Primers Expression vector Genes Immunoglobulin Antibodies Monoclonal General Medicine Molecular biology Recombinant Proteins Restriction site Plasmids |
| Zdroj: | Gene. 187:9-18 |
| ISSN: | 0378-1119 |
| Popis: | 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, αD11, and the V regions from 2E10, a scFv selected from a scFv phagemid library. |
| Databáze: | OpenAIRE |
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