Monoclonal antibody to polycyclic aromatic hydrocarbons based on a new benzo[a]pyrene immunogen

• Published in: Analytical and bioanalytical chemistry 2001-11, Vol.371 (5), p.578-585
• Main Authors: SCHARNWEBER, T, FISHER, M, SUCHANEK, M, KNOPP, D, NIESSNER, R
• Format: Article
• Language: English
• Subjects: Air Pollutants - analysis; Albumin; Analytical chemistry; Animals; Antibodies, Monoclonal - chemistry; Antigens; Biological and medical sciences; Biosensors; Biotechnology; Chemical plants; Chemistry; Chromatography, High Pressure Liquid; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Exact sciences and technology; Female; Fundamental and applied biological sciences. Psychology; Glycerin; Glycerol; Methods. Procedures. Technologies; Mice; Mice, Inbred BALB C; Miscellaneous; Monoclonal antibodies; Polycyclic aromatic hydrocarbons; Polycyclic Aromatic Hydrocarbons - analysis; Polycyclic Aromatic Hydrocarbons - chemistry; Polycyclic Aromatic Hydrocarbons - immunology; Saturated fatty acids; Sensitivity and Specificity; Various methods and equipments
• ISSN: 0937-0633; 1618-2642; 1432-1130
• DOI: 10.1007/s002160101012

Benzo[a]pyrenebutyric acid (B[a]PBA) has been synthesized and covalently coupled to bovine serum albumin to generate monoclonal antibodies (Mab). A competitive indirect enzyme-linked immunosorbent assay (ELISA) for polycyclic aromatic hydrocarbons (PAH) has been developed with Mab B[a]P-13. It was shown by testing with 21 parent PAH and 10 compounds carrying methyl, hydroxy, or butyric acid functions that the antibody had broad specificity. Highest affinity was found for four- to six-ring PAH. Different organic co-solvents were tested. No loss in sensitivity, compared with controls in PBS, were found with methanol, dimethyl sulfoxide, and glycerol at final concentrations of 5 to 10%. Further, an observation was made that a modification (fine-tuning) of the affinity and specificity of the antibodies was possible by changing the type of the added organic co-solvent. The high susceptibility of the ELISA with regard to inorganic ions might be an indication of a more hydrophilic binding pocket e.g. involving a π-cation interaction. Investigation of the effect of pH revealed that for pH between 6 to 9 there was no noticeable impairment. With an LOD as low as 30 pg per well for B[a]P the sensitivity of the ELISA is sufficient for analyses of solvent extracts of many environmental samples. As an example, the determination of a PAH sum parameter, given as B[a]P-equivalents, in crude aerosol extracts by both ELISA and HPLC revealed good correlation (r²=0.717) but approximately five-fold overestimation by the immunochemical method, obviously as a result of cross-reacting analytes.