Boronate-mediated antibody orientation: a molecular simulation-guided strategy for ultra-sensitive lateral flow immunoassays
Microchimica Acta, cilt.193, sa.9, 2026 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 193 Sayı: 9
- Basım Tarihi: 2026
- Doi Numarası: 10.1007/s00604-026-08331-w
- Dergi Adı: Microchimica Acta
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Chemical Abstracts Core, Chimica, Compendex, EMBASE, MEDLINE, Academic Search Ultimate (EBSCO), Natural Science Collection (ProQuest), Biological Science Database (ProQuest), Engineering Source (EBSCO), Health Research Premium Collection (ProQuest), Materials Science & Engineering Collection (ProQuest), Technology Collection (ProQuest)
- Anahtar Kelimeler: Antibody orientation, Boronate chemistry, Food safety, Lateral flow immunoassay, Molecular dynamics simulation, Zearalenone
- Sivas Cumhuriyet Üniversitesi Adresli: Evet
Özet
The random immobilization of antibodies on lateral flow immunoassay (LFIA) strips remains a major factor limiting assay sensitivity and consistency. To address this, we report a boronate affinity-based strategy for site-specific antibody orientation via covalent reaction between boronic acid and cis-diol groups on Fc-region glycans. Molecular dynamics simulations provided mechanistic support for this design. The boronate-antibody configuration demonstrated the most favorable binding energy (− 153.93 kJ/mol) and the fastest conformational stabilization (< 40 ns), suggesting a stabilization process involving initial electrostatic guidance followed by hydrogen-bond and covalent bond formation. Experimentally, this oriented immobilization achieved an antibody conjugation efficiency of 99.92%, a Fab exposure rate of 47.52% and significantly improved functional activity, with an affinity constant of 0.93 × 10⁸ M⁻¹, outperforming conventional passive adsorption. Applied to the detection of zearalenone (ZEN) in corn, the oriented LFIA showed a dramatic sensitivity enhancement. The limit of detection reached 0.049 µg/kg, 200-fold higher than traditional colloidal gold LFIA. The visual cut-off value was 0.5 µg/kg. Validation with spiked and naturally contaminated samples yielded recoveries of 84.7%–103.4% (CV: 7.1%–12.6%) and excellent correlation (R2 > 0.97) with LC-MS/MS. This work establishes a rational, chemistry-driven paradigm for antibody immobilization that can be generalized to enhance the performance of next-generation rapid diagnostics.