Elisa (enzyme-linked immuno-sorbent assay) kits for detection of environmental quality

New elisa kits for detection of environmental pollutants on the basis of new monoclonal antibodies

Industry development, intensive agriculture and traffic increases have a negative influence on the quality of the environment. The decreasing quality of the environment has a negative influence on the health and reproduction of mammals, including humans. Except for global procedures for environmental control (e.g. the Kyoto protocol), a challenge for modern society is to monitor and control natural and man-made harmful compounds in a given territory. Different pollutants may injure biological functions of the organisms and consequently cause problems in reproduction. Among deleterious drugs are endocrine disruptors, which can mimic or interfere with the action of endogenous estrogenic hormones. Phytoestrogens are compounds that are naturally present in edible plant material; xenoestrogens are man-made chemicals and can be present as food contaminants, which constitute a reproductive toxicology risk. Recent studies indicate that more than 15% of the European population suffers from infertility-related problems (according to the World Health Organization, 1998). For these reasons, detection of such drugs is important for human health and reproduction. The consequences of negative factors for human health manifest themselves as disorders of the immunological apparatus (such as allergy, asthma, constitutional dermatitis) and also by immunological failures in reproduction (antibodies against sperm, against the egg and also against trophoblast proteins). The present project will concentrate on the research and development of suitable detection systems based on polyclonal and monoclonal antibodies, which will test selected toxic pollutants and selected immunological diseases. SPECIFIC OBJECTIVES ARE: 1. To prepare an ELISA (Enzyme-Linked Immuno-Sorbent Assay) kit for the detection of polycyclic aromatic hydrocarbons (PAHs), especially benzpyrene. Benzpyrene is present in city atmospheres (consequence of high traffic volumes in cities). Benzpyrene has mutagenic and allergic effects. A monoclonal antibody for detection of this pollutant was prepared in the CZECH REPUBLIC (Project EUREKA OE 72). The first aim of this part of the project will be the construction of the ELISA kit. 2. To prepare monoclonal antibodies and then ELISA kits for the detection of selected endocrine disruptors. Bisphenol-A (BPA) and para-nonylphenol (NP) are endocrine disruptors. BPA is a well-known toxicant whose estrogenic activity has been known about for a long time. Recently, it was described as being released from polycarbonate flasks. NP is used in the preparation of lubrication oil additives, plasticizers and surface-active agents. It has been found in polyvinylchloride (PVC) used in the food processing and packing industries. NF ranks among alkyl phenols and accumulates in the lipids of living organisms. These drugs interfere with the normal function of endocrine systems. Recently it was found that they had a negative effect on in-vivo fertility of animals (Peknicova et al., 2002, Kyselova et al., 2003, 2004, etc.). They can probably have a similar effect on the increase in human infertility. The first aim of this part of the project is to prepare monoclonal antibodies and later to prepare kits for their detection. 1. Peknicova J., Kyselova V., Buckiova D., Boubelik M: Effect of endocrine disruptor on mammalian fertility. Application of monoclonal antibodies against sperm proteins as markers for testing sperm damage. Amer. J. Reprod. Immunol.47:311-318,2002 2. Kyselova V., Peknicova J., Buckiova D., Boubelik M.: Effect of p-nonylphenol and resveratrol on in-vivo fertility and different body parameters in outbred mice CDl, Reprod. Biol. Endocrinol. 1:30,2003 ( 3. Kyselova V., Peknicova J., Boubelik M., Buckiova D.: Body and organ weight, sperm acrosomal status and reproduction after genistein and diethylstilbestrol treatment of CD1 mice in a multigenerational study. Theriogenology, 61: 1307-1325,2004 3. To prepare an ELISA kit for the detection of autoantibodies against trophoblast proteins with a prepared monoclonal antibody as a positive control. To prepare and characterise other monoclonal antibodies for practical application. In the group of infertile females, we found autoantibodies against trophoblast proteins as a consequence of unfavourable environmental conditions. The first antibody was prepared with VIDIA COOPERATION (Centre for molecular and gene biotechnology (LNB030), while other antibodies will be prepared, characterised and used in the ELISA kit. Keywords: ELISA (Enzyme-Linked Immuno-Sorbent Assay) kits, monoclonal antibody, pollutants.
Project ID: 
3 557
Start date: 
Project Duration: 
Project costs: 
440 000.00€
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