Chemistry: analytical and immunological testing – Involving producing or treating antigen or hapten
Patent
1996-04-26
1998-11-03
Housel, James C.
Chemistry: analytical and immunological testing
Involving producing or treating antigen or hapten
435 792, 435 793, 436547, 436548, 5303889, G01N 33531, G01N 3353, C07K 1600
Patent
active
058307702
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to novel generic antibodies that are specific to groups of organophosphates, particularly to a class of organophosphate pesticides in common usage. Further provided are hapten compounds and methods for using them in raising these antibodies, test kits containing them and methods for their use.
A number of immunoassays are available for the detection of various biologically important compounds. Such compounds include the important organophosphate (OP) class of pesticides (see eg. WO 91/00294). These immunoassays are relatively sensitive, but are directed against individual OP molecules and thus several separate assays are required to effectively screen a material for OP presence.
A group of these organophosphate pesticides that is commonly used for foodstuff, eg. grain, protection comprises the compounds methacrifos, pirimphos methyl, fenitrothion, etrimfos, propetamphos, chlopyrifos methyl and dichlorvos, and screening a material for the presence of any of these potentially toxic compounds conventionally requires use of seven separate agents. While these compounds share some chemical features with each other, including that of the OP group itself, other commonly used pesticides are distinguished by variant OP groups. In malathion for example a (CH.sub.3 O).sub.2 P(S)--S-- group is present as compared to the (CH.sub.3 O).sub.2 P(S)--O-- group of the compounds referred to above.
The present inventors have now provided a novel strategy for raising antibodies such that they are specifically directed to bind with two or more of these specified pesticidal organophosphorous compounds and further provide methods for using these and kits containing them, such that materials can be screened for presence of a member of a selected group with a single antibody reagent. Still further they have provided organic haptens and their conjugates which may be used specifically to raise these antibodies, or antisera containing them, as either monoclonal or polyclonal antibodies. Particularly provided are antibodies capable of specific binding with two or more, more preferably three or more, and most preferably all of the compounds methacrifos, fenitrothion, etrimfos, propetamphos, dichlorvos and optionally one or more of malathion, dimethoate, chlorfenvinphos, chlopyrifos methyl, tetrachlorvinphos and pirimiphos-methyl, wherein the relative binding affinity with each of these bound compounds is such that a single strength antibody reagent can be used to bind them. Preferably the OPs bound are capable of displacing percentages of bound hapten from the antibody at a concentration of 100 .mu.g/ml within a factor of 20, more preferably of 5, and more preferably 2 of each other. Also provided are haptens and hapten-protein conjugates which evoke production of such antibodies when adminstered to animals.
The concept that one generic antibody or antisera might be raised to be capable of detecting a number of organophosphates has been considered on one occasion previously by Suedi and Heesham Kiel: (1988) Milchwirt Forsch 40 179-202, but this involved use of a number of model pesticide molecules to provide a number of protein conjugates. Furthermore the antibody reagents obtained were not capable of use in reliable screening.
In a first aspect of the present invention there is provided a hapten protein conjugate capable of stimulating production of antibodies which have specific binding affinity for compounds including the chemical moiety I: characterised in that the antibody binds two or more, preferably three or more, and most preferably all of the compounds methacrifos, fenitrothion, propetamphos, dichlorvos, dimethoate, malathion, chlorfenvinphos and etrimfos; preferably also binding to chlopyrifos methyl, tetrachlorvinphos and pirimiphos-methyl.
Most preferably the conjugates evoke production of antibodies targeted at moiety (I) where R is methyl, and preferably where A is O, wherein cross-reactivity with other compounds relative to those having this moiety is low enough to enable ready identification
REFERENCES:
patent: 5283180 (1994-02-01), Zomer et al.
patent: 5413915 (1995-05-01), Case et al.
patent: 5589624 (1996-12-01), Ulbrich et al.
Banks John Nicholas
Chaudhry Mohamed Qasim
Matthews Wendy Alison
Housel James C.
Swartz Rodney P.
The Minister of Agriculture Fisheries and Food in her Britannic
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