Efficient Discovery of Immunoglobulin Genes for Detecting Organophosphorus Pesticides Using a Reverse Virtual...

Area of Science:

• Bioanalytical Chemistry
• Immunology
• Computational Biology

Background:

• Identifying genes for hapten-specific recombinant antibodies (rAbs) is challenging.
• Hapten-specific rAbs are vital for tracing small molecules in bioanalysis.

Purpose of the Study:

• To establish an efficient reverse virtual screening pipeline for discovering variable region (VR) sequences of hapten-specific rAbs.
• To identify and characterize a recombinant antibody targeting parathion and related pesticides.

Main Methods:

• Constructed homology models of antibody fragment variable (Fv) regions using diverse VR sequences.
• Employed energy scoring and molecular interaction analysis for virtual screening.
• Validated antibody affinity and specificity using surface plasmon resonance and ELISA assays.
• Elucidated the recognition mechanism via molecular dynamics simulations.

Main Results:

• Successfully identified and validated a recombinant antibody (rAb-Model 6) with high affinity for parathion.
• Demonstrated broad specificity for parathion-methyl and fenitrothion (Kd: 3.84 × 10⁻⁹–1.07 × 10⁻⁸ M).
• rAb-Model 6 showed high sensitivity for three EU-banned organophosphorus pesticides (IC50: 4.94–9.70 ng/mL).

Conclusions:

• The developed in silico pipeline is efficient and cost-effective for identifying anti-hapten immunoglobulin genes.
• The study provides a promising tool for the simultaneous monitoring of parathion, parathion-methyl, and fenitrothion.
• This strategy aids in discovering anti-parathion antibody genes and developing new bioanalytical tools.