Chemistry: molecular biology and microbiology – Carrier-bound or immobilized enzyme or microbial cell;...
Patent
1987-11-24
1991-10-29
Russel, Jeffrey E.
Chemistry: molecular biology and microbiology
Carrier-bound or immobilized enzyme or microbial cell;...
260403, 435192, 435207, 436545, 436546, 530345, 530402, 530403, 530404, 530406, 530408, 530409, 530410, 558169, 558172, C12N 1100, C07K 1700, C07F 909, G01N 33532
Patent
active
050616269
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to novel antigens capable of producing antibodies to Platlet Activating Factor (PAF), novel PAF analoques labelled to enable quantitative assay, intermediates for the production of novel PAF antigens and methods for the preparation of said antigens, and methods of immunoassay of PAF in biological fluid using said labelled analogues and/or labelled PAF-antibodies.
BACKGROUND
Platelet Activating Factor (PAF), 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine, represents a recently defined example of a class of biologically-active lipids active in the subnanomolar range and possessing a wide spectrum of pathophysiological effects. PAF promotes life-threatening anaphylactic reactions in animals and is suspected of mediating a range of allergic and inflammatory reactions in man. For example, PAF may be important in conditions such as asthma, adult respiratory distress syndrome and shock reactions. However, despite the increasing catalogue of conditions in which PAF maybe involved, greater insights into its role in health and disease ar hampered because precise and specific methods for its measurement are lacking. The capacity of PAF to aggregate platelets does not provide a suitable basis for strictly quantitative assay.
It would be desirable to develop an immunoassay for quantitative determination of PAF levels in blood serum. However, it has been found that PAF itself is insufficiently antigenic to produce the necessary PAF-antibodies needed for such an immunoassay.
Novel synthetic PAF analogues have now been found which are sufficiently antigenic to produce PAF-antibodies and a method suitable for the immunoassay of PAF levels in biological fluids has been developed.
THE INVENTION
Accordingly the invention provides novel compounds of general formula (I) ##STR2## wherein: (1) R.sup.1 is a C.sub.2 to C.sub.25 alkylene or alkenylene linking group substituted by radioactive iodine; linking group optionally substituted by tritium or radioactive iodine; carboxy, isothiocyanato, N--C.sub.1 to C.sub.6 alkylamino, N,N-di(C.sub.1 to C.sub.6 alkyl)amino, hydroxy and mercapto; and --NR.sup.6 --, --COO--, --OCO--, --CONR.sup.6 --, --NR.sup.6 CO--, --NH--CS--NH--and --S--S--wherein R.sup.6 is selected from hydrogen and C.sub.1 to C.sub.6 alkyl; and lipid groups and derivatives thereof of molecular weight of at least 2000; and alkyl; and mixtures of the compound of formula (I) and its enantiomer.
In one embodiment the invention provides antigenic PAF analogues of general formula (I) wherein: --CONR.sup.6 --, --NR.sup.6 CO--and --S--S--wherein R.sup.6 is selected from hydrogen and C.sub.1 to C.sub.6 alkyl; and carbohydrate and lipid groups and derivatives thereof of molecular weight of at least 2000 which are capable of eliciting an antigenic response; and alkyl.
In the antigenic PAF analogues of the invention of general formula (I):
Preferred R.sup.1 include straight chain C.sub.4 to C.sub.16 alkylene. More preferred R.sup.1 include straight chain C.sub.4 to C.sub.8 alkylene. For convenience R.sup.1 is often chosen from pentylene and hexylene.
Preferred A include --NR.sup.6 --, --COO--, --OCO--, --CONR.sup.6 --and R.sup.6 CO--and preferred R.sup.6 include hydrogen and methyl. More preferred A include --NR.sup.6 --and --OCO--.
Preferred B include monofunctional and polyfunctional protein, peptide, carbohydrate and lipid groups of molecular weight at least 5000 and capable of eliciting an antigenic response. More preferred B include monofunctional and polyfunctional groups of molecular weight at least 10,000. Examples of suitable B include Bovine Serum Albumin (BSA), ovalbumin, Porcine Thyroglobulin (PTG), Bovine Thyroglobulin (BTG), keyhole limpet hemocyanin, bacterial cell walls, synthetic polypeptides such as polylysine, poke weed mitagen (PWM), phytohaemoglutinin (PHA), muramyl dipeptidase and lipopolysaccharides.
Preferred R.sup.2 to R.sup.5 include C.sub.1 to C.sub.3 alkyl, and especially methyl.
In another embodiment the invention provides labelled PAF analogues of g
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Baldo Brian A.
Redmond John W.
Kim Kay
Russel Jeffrey E.
University of Sydney
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