Production of monoclonal antibodies against fullerene C sub(60) and development of a fullerene enzyme immunoassay

• Published in: Analyst (London) 2012-01, Vol.137 (1), p.98-105
• Main Authors: Hendrickson, O, Fedyunina, N, Zherdev, A, Solopova, O, Sveshnikov, P, Dzantiev, B
• Format: Article
• Language: English
• Subjects: Antibodies; Assaying; Biological; Buckminsterfullerene; ELISA; Enzymes; Fullerenes; Immunoassay
• ISSN: 0003-2654
• DOI: 10.1039/c1an15745k

The aim of the present study was to produce monoclonal anti-fullerene C sub(60) antibodies and to develop the enzyme immunoassay for the detection in the first use of free fullerene C sub(60) both in solutions and in multicomponent biological probes. The immunization of mice with the conjugate of fullerene C sub(60) carboxylic derivative with thyroglobulin synthesized by carbodiimide activation led to the production of eight clones of anti-fullerene antibodies. The specificity of the antibody-fullerene binding was confirmed. Indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed for the determination of water-soluble protein-conjugated fullerene, the fullerene aminocaproic acid, fullerenol and for pristine fullerene in solution. To solubilize extremely hydrophobic free fullerene C sub(60) a specially selected water-organic mixture compatible with immunoassay was proposed. The detection limit of free fullerene C sub(60) in solution was 2 mu g L super(-1). Fullerene C sub(60) was also detected by ELISA in organ homogenates of rats intraperitoneally or intragastrically administered with fullerene. To reduce the influence of biomatrices on the assay results a technique was developed for the biological sample pretreatment by the extraction of C sub(60) from bioprobe by toluene followed by the evaporation of toluene and dissolution of the fullerene-containing extract in the selected water-organic media. The ELISA procedure in the first use allowed the detection of fullerene C sub(60) in different tissues.