Development of a novel scintillation proximity radiommunoassay for platelet-activating factor measurement: Comparison with bioassay and GD/MS techniques

Journal Article · Mon Jan 01 04:00:00 EST 1990 · Life Sciences; (USA)

DOI:https://doi.org/10.1016/0024-3205(90)90460-9· OSTI ID:6705207

Sagatani, Junko; Saito, Kunihiko ^[1]; Lee, D Y; Hughes, K T ^[2]

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1. Kansia Medical School, Osaka (Japan)
2. Amersham International plc., Cardiff (England)

A novel, facile and sensitive scintillation proximity radioimmunoassay (SPRIA) for quantitation of PAF has been developed. No separation of antibody bound ({sup 3}H)PAF from free ({sup 3}H)PAF is required as the assay employs protein A - coated fluomicrospheres (beads containing scintillant). The assay system was suitable for the quantitation of 0.03 to 2 pmol of 1-hexadecyl-2-acetyl-sn-glycero-3-phosphocholine. The cross-reactivity was high with 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine but was very low with PAF analogs such as 1-alkyl- and 1-acyl-2-lyso-sn-glycero-3-phosphocholine, 1-acyl-2-acetyl-sn-glycero-3-phosphocholine, and 1-alk-1{prime}-enyl-2-acetyl-sn-glycero-3-phosphocholine, and 1-alk-1{prime}-enyl-2-acetyl-sn-glycero-3-phosphocholine. The specificity of SPRIA was higher than that of bioassay (platelet degranulation assay). PAF receptor antagonists (L-652,731, WEB2086, and FR900452) at up to 10 nmol per tube had no affect on the SPRIA. These observations indicate that the specificity of the PAF antibody is quite different from that of the platelet receptor. The values obtained using SPRIA for the measurement of PAF produced in polymorphonuclear leukocytes with stimuli are comparable to those obtained by SIM/GC/MS analysis.

• OSTI ID: 6705207
• Journal Information: Life Sciences; (USA), Journal Name: Life Sciences; (USA) Vol. 46:20; ISSN 0024-3205; ISSN LIFSA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANTIBODIES
BIOASSAY
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BLOOD COAGULATION FACTORS
BODY FLUIDS
CHROMATOGRAPHY
COAGULANTS
COMPARATIVE EVALUATIONS
DIAGNOSTIC TECHNIQUES
DRUGS
GAS CHROMATOGRAPHY
HEMATOLOGIC AGENTS
HYDROGEN COMPOUNDS
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
LEUKOCYTES
MASS SPECTROSCOPY
MATERIALS
ORGANIC COMPOUNDS
PROTEINS
RADIOASSAY
RADIOIMMUNOASSAY
RADIOIMMUNODETECTION
RADIOIMMUNOLOGY
SEPARATION PROCESSES
SPECIFICITY
SPECTROSCOPY
TRACER TECHNIQUES
TRITIUM COMPOUNDS