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Titolo:
High-throughput screening of surface displayed gene products
Autore:
Walter, G; Konthur, Z; Lehrach, H;
Indirizzi:
Biochard AS, N-0281 Oslo, Norway Biochard AS Oslo Norway N-0281Biochard AS, N-0281 Oslo, Norway
Titolo Testata:
COMBINATORIAL CHEMISTRY & HIGH THROUGHPUT SCREENING
fascicolo: 2, volume: 4, anno: 2001,
pagine: 193 - 205
SICI:
1386-2073(200104)4:2<193:HSOSDG>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
LINKED-IMMUNOSORBENT-ASSAY; PROTEIN EXPRESSION; SOMATIC HYPERMUTATION; AFFINITY MATURATION; CDNA LIBRARY; PHAGE; ANTIBODY; SELECTION; SEQUENCE; CLONING;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
42
Recensione:
Indirizzi per estratti:
Indirizzo: Walter, G Biochard AS, Nedre Skogvei 14, N-0281 Oslo, Norway Biochard AS Nedre Skogvei 14 Oslo Norway N-0281 81 Oslo, Norway
Citazione:
G. Walter et al., "High-throughput screening of surface displayed gene products", COMB CHEM H, 4(2), 2001, pp. 193-205

Abstract

With the human genome project approaching completion, there is a growing interest in functional analysis of gene products. The characterization of large numbers of proteins, their expression patterns and in vivo localisations, demands the use of automated technology that maintains a logistic link to the encoding genes. As a complementary approach, phage display is used for recombinant protein expression and the selection of interacting (binding)molecules. Cloning of libraries in filamentous bacteriophage or phagemid vectors provides a physical link between the expressed protein and its encoding DNA sequence. High-throughput technology for automated library handlingand phage display selection has been developed using picking-spotting robots and a module for pin-based magnetic particle handling. This system enables simultaneous interaction screening of libraries and the selection of binders to different target molecules at high throughput. Target molecules areeither displayed on high-density filter membranes (protein filters) or tag-bound to magnetic particles and can be handled as native ligands. Binding activity is confirmed by magnetic particle ELISA in the microtitre format. The whole procedure from immobilisation of target molecules to confirmed clones of binders is automatable. Using this technology, we have selected human scFv antibody fragments against expression products of human cDNA libraries.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/07/20 alle ore 17:30:44