Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
A METHOD FOR COMBINED IMMUNOAFFINITY PURIFICATION AND ASSAY OF HIV-1 REVERSE-TRANSCRIPTASE ACTIVITY USEFUL FOR CRUDE SAMPLES
Autore:
LENNERSTRAND J; RYTTING AS; ORVELL C; GRONOWITZ JS; KALLANDER CFR;
Indirizzi:
UNIV UPPSALA,CTR BIOMED,DEPT MED GENET,RES UNIT REPLICAT ENZYMOL,BOX 584 S-75123 UPPSALA SWEDEN SANGTEC MED AB S-16120 BROMMA SWEDEN HUDDINGE HOSP S-14186 HUDDINGE SWEDEN
Titolo Testata:
Analytical biochemistry
fascicolo: 2, volume: 235, anno: 1996,
pagine: 141 - 152
SICI:
0003-2697(1996)235:2<141:AMFCIP>2.0.ZU;2-D
Fonte:
ISI
Lingua:
ENG
Soggetto:
HUMAN-IMMUNODEFICIENCY-VIRUS; MONOCLONAL-ANTIBODIES; CAPTURE ASSAY; RNASE-H; TYPE-1; POLYMERASE; PROCESSIVITY; HETERODIMER; TEMPLATES; ENZYMES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
33
Recensione:
Indirizzi per estratti:
Citazione:
J. Lennerstrand et al., "A METHOD FOR COMBINED IMMUNOAFFINITY PURIFICATION AND ASSAY OF HIV-1 REVERSE-TRANSCRIPTASE ACTIVITY USEFUL FOR CRUDE SAMPLES", Analytical biochemistry, 235(2), 1996, pp. 141-152

Abstract

Detection of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) activity in crude specimens was greatly enhanced using a novel capture RT assay, Eighteen different monoclonal antibodies (Mabs) raised against purified HIV-1 RT were tested for their ability to bind to HIV-1 RT without affecting its activity. The anti-HIV-1 RT Mabs were immobilized on plastic macrobeads and used as solid carriers inthe capture RT assay. The assay system first involved RT's adherence to the immobilized Mabs, Nonspecific enzymes and other impurities wereremoved by a simple wash after which the RT reaction mixture was added, Substrate and product were finally separated by a wash of the beads, Practically all radioactivity incorporated into DNA (>98%) was recovered on the bead, The Michaelis-Menten constants and the saturation velocity values for the nucleotide substrate were similar for free and immobilized RT. The reaction mechanism for the immobilized RT is discussed, When comparing the function of this assay with more conventional soluble RT assays for samples consisting of recombinant HIV-1 RT mixedwith an extract of peripheral blood lymphocytes (PBL), an almost 100-fold higher sensitivity was found, The capture RT assay had the capacity to recover approximately 80% of the RT activity added to an extractof 1 X 10(7) PBL cells/ml. A strong correlation (r = 0.947) between the results obtained with this assay and a HIV-1 p24 enzyme-linked immunosorbent assay was found, when samples from a collection of 16 HIV strains propagated in cell culture were analyzed. (C) 1996 Academic Press, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 13:56:00