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Cited by 87 publications
(101 citation statements)
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“…Newly developed ways of building antibody libraries, especially in the scFv format, and new methods for selection from those libraries, including phage display, yeast twohybrid and mammalian two-hybrid, solved some of the problems by facing the development of intrabodies as therapeutic agents. The development in the selection methods led to the definition of an intrabody consensus sequence, which improved expression properties in vivo (Marasco, 1997b;Tse et al, 2002;Visintin et al, 2002;Stocks, 2005).…”
Section: Intracellular Antibodies (Intrabodies)mentioning
confidence: 99%
“…Newly developed ways of building antibody libraries, especially in the scFv format, and new methods for selection from those libraries, including phage display, yeast twohybrid and mammalian two-hybrid, solved some of the problems by facing the development of intrabodies as therapeutic agents. The development in the selection methods led to the definition of an intrabody consensus sequence, which improved expression properties in vivo (Marasco, 1997b;Tse et al, 2002;Visintin et al, 2002;Stocks, 2005).…”
Section: Intracellular Antibodies (Intrabodies)mentioning
confidence: 99%
“…Expression of scFvs in yeast is an attractive alternative since established methodologies are available that allow screening of diverse libraries. Furthermore, functionality of antibody fragments in the cytoplasm and nucleus of lower eukaryotic cells appears to be predictive for their behavior upon expression in the intracellular milieu of mammalian cells (Visintin et al, 1999;De Jaeger et al, 2000;Wo¨rn et al, 2000;Tse et al, 2002). After enrichment of phage-displayed, EGFR-specific antibody fragments from a combinatorial library and transfer into a yeast two-hybrid screen, we could isolate a panel of scFvs that are active in the cytosol and bind strongly to the intracellular domain of human EGFR.…”
Section: Discussionmentioning
confidence: 99%
“…Indeed, direct yeast two-hybrid screening of a portion of the unselected anti-EGFR scFv library (approximately 10 5 independent plasmids) did not result in the identification of functional EGFR-specific intrabodies (data not shown). Owing to limitations in transformation efficiency, quantitative transfer of combinatorial scFv libraries of more than approximately 10 6 independent clones into a yeast screening system is not practical (Tse et al, 2002). Therefore, EGFR-binding antibody fragments in our initial phage display library were enriched by three rounds of biopanning, reducing the complexity of the library to a value below 10 6 , which proved suitable for subsequent expression in yeast.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…We have used intra-cellular antibody capture technology (Tse et al, 2002; to isolate an anti-LMO2 scFv (Anti-LMO2 clone R3, ALR3) following the steps indicated (Figure 1). Initially a diverse phage scFv library was screened in vitro using an LMO2 fusion protein and 1.9 Â 10 5 binders were isolated ( Figure 1a).…”
mentioning
confidence: 99%