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Titolo:
Peptide and small molecule microarray for high throughput cell adhesion and functional assays
Autore:
Falsey, JR; Renil, M; Park, S; Li, SJ; Lam, KS;
Indirizzi:
Univ Calif Davis, Ctr Canc, Div Hematol Oncol, Sacramento, CA 95817 USA Univ Calif Davis Sacramento CA USA 95817 Oncol, Sacramento, CA 95817 USA Univ Calif Davis, Dept Internal Med, Sacramento, CA 95817 USA Univ Calif Davis Sacramento CA USA 95817 al Med, Sacramento, CA 95817 USA
Titolo Testata:
BIOCONJUGATE CHEMISTRY
fascicolo: 3, volume: 12, anno: 2001,
pagine: 346 - 353
SICI:
1043-1802(200105/06)12:3<346:PASMMF>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN-TYROSINE KINASES; COMBINATORIAL LIBRARY METHOD; PARALLEL CHEMICAL SYNTHESIS; DRUG DISCOVERY; ORTHOGONAL LIGATION; BUILDING-BLOCKS; SUBSTRATE; IDENTIFICATION; SPECIFICITY; SUPPORTS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
42
Recensione:
Indirizzi per estratti:
Indirizzo: Lam, KS Univ Calif Davis, Ctr Canc, Div Hematol Oncol, 4501 X St, Sacramento, CA 95817 USA Univ Calif Davis 4501 X St Sacramento CA USA 95817 o, CA 95817 USA
Citazione:
J.R. Falsey et al., "Peptide and small molecule microarray for high throughput cell adhesion and functional assays", BIOCONJ CHE, 12(3), 2001, pp. 346-353

Abstract

A novel class of chemical microchips consisting of glass microscope slideswas prepared for the covalent attachment of small molecule ligands and peptides through site-specific oxime bond or thiazolidine ring ligation reaction. Commercially available microscope slides were thoroughly cleaned and derivatized with (3-aminopropyl)triethoxysilane (APTES). The amino slides were then converted to glyoxylyl derivatives via two different routes: (1) coupling of Fmoc-Ser followed by deprotection and oxidation, or (2) coupling with protected glyoxylic acid and final deprotection with HCl. Biotin or peptide ligands derivatized at the carboxyl terminus with a 4,7,10-trioxa-1,13-tridecanediamine succinimic acid linker and an amino-oxy group or a 1,2-amino-thiol group (e.g., cysteine with a free N-alpha-amino group) mere printed onto these slides using a DNA microarray spotter. After chemical ligation, the microarray of immobilized ligands was analyzed with three different biological assays: (1) protein-binding assay with fluorescence detection, (2) functional phosphorylation assay using [gamma P-33]-ATP and specific protein kinase to label peptide substrate spots, and (3) adhesion assay with intact cells. In the cell adhesion assay, not only can we determine the binding specificity of the peptide against different cell lines, we can also determine functional cell signaling of attached cells using immunofluorescence techniques in situ on the microchip. This chemical microchip system enables us to rapidly analyze the functional properties of numerous ligands thatwe have identified from the "one-bead one-compound" combinatorial Library method.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 09/07/20 alle ore 00:47:12