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A sensitive and specific radioimmunoassay for platelet-activating factor

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Lipids

Abstract

A platelet-activating factor (PAF) analog with a reactive ω-aldehyde group at thesn-1 position was synthesized. The hapten-thyroglobulin conjugate was used to immunize rabbits to produce specific antibodies to PAF. The purified immunoglobulin G (IgG) fraction was found to bind stereo-specifically to tritiated PAF and to crossreact minimally with lysoPAF, plasmalogens, and other phospholipids. The radioimmunoassay detected as little as 20 pg of PAF per assay tube and was used to explore agonist-induced synthesis of PAF in rabbit neutrophils. Calcium ionophore A23187 at 1 μM induced PAF synthesis peaking at 2 min and reaching basal levels after 5 min.N-Formyl-Met-Leu-Phe (FMLP) at 0.1 μM also stimulated rapid synthesis and degradation of PAF with a peak at 5 min. Both A23187 and FMLP stimulated PAF synthesis in a dose-dependent manner. The radioimmunoassay should be applicable to the quantitation of PAF in biological samples.

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Abbreviations

CPL:

choline plasmalogen

FMLP:

N-formyl-methionylleucyl-phenylalanine

IC50 :

50% maximal binding

IgG:

immunoglobulin G

PC:

phosphatidylcholine

PE:

phosphatidylethanolamine

PI:

phosphatidylinositol

PS:

phosphatidylserine

IysoPAF:

lyso plateletactivating factor, 1-O-alkyl-sn-glycero-3-phosphocholine

PAF:

plateletactivating factor, 1-O-alkyl-2-sn-glycero-3-phosphocholine

RIA:

radioimmunoassay

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Authors and Affiliations

  1. Division of Medicinal Chemistry and Pharmaceutics, College of Pharmacy, University of Kentucky, 40536-0082, Lexington, Kentucky

    Chang-jin Wang & Hsin-Hsiung Tai

Authors
  1. Chang-jin Wang
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  2. Hsin-Hsiung Tai
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Wang, Cj., Tai, HH. A sensitive and specific radioimmunoassay for platelet-activating factor. Lipids 27, 206–208 (1992). https://doi.org/10.1007/BF02536180

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  • DOI: https://doi.org/10.1007/BF02536180

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