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Titolo:
APPLICATION OF AN ALPHA-SIDE-CHAIN LENGTH-SPECIFIC MONOCLONAL-ANTIBODY TO IMMUNOAFFINITY PURIFICATION AND ENZYME-IMMUNOASSAY OF 2,3-DINOR-6-KETO-PROSTAGLANDIN F1-ALPHA FROM HUMAN URINE
Autore:
HIRAMATSU M; HAYASHI Y; YAMAMOTO S; HAYASHI A; SAWADA M; HAMANAKA N; YAMASHITA K;
Indirizzi:
TOKUSHIMA BUNRI UNIV,SCH MED,DEPT BIOCHEM,KURAMOTO CHO TOKUSHIMA 770 JAPAN TOKUSHIMA BUNRI UNIV,SCH MED,DEPT BIOCHEM TOKUSHIMA 770 JAPAN NIPPON KAYAKU CO LTD,PHARMACEUT GRP,RES LABS,KITA KU TOKYO 115 JAPAN ONO PHARMACEUT CO LTD,DIAGNOST RES LAB OSAKA 618 JAPAN ONO PHARMACEUT RES CO LTD,MONASE RES INST OSAKA 618 JAPAN
Titolo Testata:
Prostaglandins, leukotrienes and essential fatty acids
fascicolo: 2, volume: 50, anno: 1994,
pagine: 69 - 79
SICI:
0952-3278(1994)50:2<69:AOAALM>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
CHROMATOGRAPHY-MASS-SPECTROMETRY; PROSTACYCLIN METABOLITES; QUANTITATIVE-ANALYSIS; LINKED IMMUNOASSAY; THROMBOXANE A2; METHYL-ESTERS; SOLID-PHASE; RAT URINE; EXTRACTION; EXCRETION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
48
Recensione:
Indirizzi per estratti:
Citazione:
M. Hiramatsu et al., "APPLICATION OF AN ALPHA-SIDE-CHAIN LENGTH-SPECIFIC MONOCLONAL-ANTIBODY TO IMMUNOAFFINITY PURIFICATION AND ENZYME-IMMUNOASSAY OF 2,3-DINOR-6-KETO-PROSTAGLANDIN F1-ALPHA FROM HUMAN URINE", Prostaglandins, leukotrienes and essential fatty acids, 50(2), 1994, pp. 69-79

Abstract

2,3-Dinor-6-keto-prostaglandin F-1 alpha (2,3-dinor-6-keto-PGF(1 alpha)) is a major urinary metabolite of PGI(2) (prostacyclin) and one of the most reliable parameters of PGI(2) production in vivo. A mouse wasimmunized with 2,3-dinor-6-keto-20-carboxy-PGF(1 alpha) conjugated tobovine serum albumin for preparation of a monoclonal antibody which recognized the difference in the a-sidechain length of 6-keto-PGF(1 alpha) and its 2,3-dinor-metabolite. A sensitive and specific enzyme immunoassay was developed by the solid-phase competition method with 2,3-dinor-6-keto-20-carboxy-PGF(1 alpha) labeled with peroxidase protein. The detection range of the assay was 14-1200 fmol (IC50 = 120 fmol). The cross-reactivities of the antibody with 6-keto-PGF(1 alpha), 6,15-diketo-13,14-dihydro-PGF(1 alpha), and other arachidonate metabolites were less than 0.01%. An immunoaffinity column was prepared by coupling the anti-2,3-dinor-6-keto-PGF(1 alpha) antibody to BrCN-activated Sepharose 4B. Human urine was applied to an octadecylsilyl silica catridge, and the extract was applied to the immunoaffinity column. This procedure allowed an efficient separation of 2,3-dinor-6-keto-PGF(1 alpha),from unidentified urinary substances which interfered with immunoassay. Validity of the results obtained by the enzyme immunoassay was confirmed by CC/MS employing selected ion monitoring for quantification.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 07/07/20 alle ore 04:51:22