High-affinity uranyl-specific antibodies suitable for cellular imaging

Area of Science:

• Immunology
• Toxicology
• Analytical Chemistry

Background:

• Monoclonal antibodies (mAbs) are effective tools for studying protein-metal interactions.
• Previous research has identified specific antibodies for chelated metal ions, including uranyl.

Purpose of the Study:

• To generate novel mAbs with high specificity for uranyl.
• To investigate the cross-reactivity of these mAbs with various uranyl complexes.
• To explore the application of these mAbs in toxicological studies, particularly in visualizing uranyl within cells.

Main Methods:

• Immunization of mice with UO2(2+)-DCP-BSA to produce mAbs.
• Counter-screening using UO2(2+)-DCP-casein to select specific antibodies.
• Competitive binding assays with various metal ions to assess cross-reactivity.
• Fluorescence-activated cell sorting (FACS) and metal-immunostaining for cellular uranyl detection.

Main Results:

• Two highly specific anti-uranyl mAbs, U04S and U08S, were selected with dissociation constants (KD) in the picomolar range.
• Competitive assays indicated cross-reactivity with different chelating groups in the uranyl equatorial plane.
• One mAb successfully visualized intracellular uranyl accumulation in human kidney cells (HK2) using FACS and metal-immunostaining.

Conclusions:

• The developed mAbs (U04S and U08S) exhibit high specificity and affinity for uranyl.
• These mAbs demonstrate potential for detecting and visualizing uranyl in complex biological samples.
• The findings support the application of these mAbs in toxicology for assessing uranyl exposure and cellular interactions.