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Development and application of mabs-based luminescent-elisas and flow immunoassays for the detection of organochlorine pesticide residues in food

Objective


Foreseen Results

A successfull accomplishment of this research would provide to Control Authorities the basis for a new effective approach to foods and environmental monitoring of pesticides and then to a better protection of consumers health, and moreover an important contribution to the knowledge about the techniques of haptens synthesis and MAbs production, as well as their application in various research fields.
The widespread use, past and present, of a great variety of pesticides, some of them with long persistence as metabolites or degradation products, led to their presence in all compartments of the environmnent, foods and agricultural products included. Moreover, these products could be treated both on field and during storage, increasing the possibility for the accumulation of toxic residues. The recently reported data on the several and serious adverse effects on health caused by long-time, low-level exposure to toxic agrochemicals have increased worldwide the concern over pesticides residues in foods and on the need for methods with suitable analytical performance to carry out large-scale monitoring programmes on agrochemicals contaminations, i.e. able to detect in rapid, sensitive and cost-effective way low levels of pesticides residues content.
GC, or HPLC for pesticides lacking the thermal stability necessary for GC, are generally regarded as the best techniques, nevertheless they involve laborious extraction, cleanup, derivatization as well as concentration procedures to obtain the desired sensitivity. Although immunochemical technology cannot replace these techniques immunoassays are being demonstrated as an effective alternative or complementary method to the traditional ones for the routine analysis of pesticides because they are sensitive, selective, rapid and inexpensive, especially when a high sample throughput or on-side screening analysis is required.
End-point detection methods based on enzyme-mediated formation of luminescent products are another technique that is undergoing active development in many analytical fields, representing itself an amplification step increasing the sensitivity of the assay.
Among the several kinds of pesticides employed in agriculture the organochlorine ones display long persistance in the environment and, although the use is forbidden from long time, an important presence in the various components of the food chain, as well as recognized toxicity. The aim of this project is to develop a set of ELISA and flow immunoassays for the determination of organochlorous pesticides belonging to the DDT and Endosulfan families, metabolites included. The immunoassays will be based on two main innovative points: the use of Monoclonal antibodies raised against the specific antigens and the introduction, as end-point detection systems, of chemiluminescent reagents.
The immunoassays will be developed starting from the synthesis of suitable haptens, through the production and characterization of MAbs, the optimization of the ELISA format and its adaptation as chemiluminescent immunoassay. The same immunoreagents will be used to develop a flow luminescent assay, to be used as disposable reactor. Validation of the assays by comparison with results obtained using the established chromatographic techniques and the assemble of the immunoreagents as a commercial Kit are also objectives of this project, the last one to perform in close collaboration with private firms involved in the development and commercialization of immunoKits.

Coordinator

UNIVERSITY OF BOLOGNA
Address
Via San Donato 15
40127 Bologna
Italy

Participants (3)

INSTITUTE OF CHEMICAL TECHNOLOGY, PRAGUE
Czechia
Address
3,Technicka 1905/5
166 28 Praha 6
MOSCOW STATE UNIVERSITY
Russia
Address
Leninskie Gory
119 899 Moscow
Universidad Politecnica de Valencia
Spain
Address
S/n,camino De Vera
46022 Valencia