Development of a Method for Pb Functionalization and Preparing Immunogenic Complex

Antibody based methods have numerous advantages compared to other detection methods.however, heavy metals cannot stimulate the immune antibody response; it is the main obstacle forpreparation of the antibodies that used in detection methods of the metals. the production of theimmunogenic pb complexes by using cost benefit linkers of edta and dtpa were investigated inthis study.bsa molecules were functionalized using edta and dtpa linkers in variousbsa/linker ratio, ph, incubation times and buffers. the complexes were formed after addition ofthe pb to the [bsa-linker]. different concentrations of the glutaraldehyde were mixed with theformed complexes and shaken at room temperature for 12 hours. the prepared conjugates weredialyzed in phosphate buffer saline for 72 hours. stimulation of the mice antibody responses againstthe prepared [pb-linker-bsa] complex were evaluated using elisa. in optimized condition, thedtpa linker coupled pb to each bsa molecules 2 times higher than edta. the optimum ph was9.6 in edta binding to bsa; also, edta molarity was 25 times higher than pb molarity in [pb-edta-bsa] complexes. these conditions for dtpa conjugation to pb and bsa are ph 7.4-9.6and 9.6 respectively. also, optimum molarity of dtpa was 4 times higher than pb in [pb-dtpabsa]complexes. also, the higher antibody responses against pb were stimulated by immunization with an antigenic complex that has more solubility and pb coupled to each albumin molecules. inconclusion, in optimized conditions, dtpa more efficient than edta for synthesizes of theimmunogenic pb complex.