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Engineering Antiviral Agents via Surface Plasmon Resonance

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High-affinity VNARs targeting human hemoglobin: Screening, stability and binding analysis.

Wen-Hui Lei¹, Zu-Ying Liu¹, Xiao-Xiao Xie¹

¹College of Ocean Food and Biological Engineering, Jimei University, Xiamen 361021, China.

International Journal of Biological Macromolecules

|January 25, 2025

Summary

This summary is machine-generated.

Researchers developed novel shark-derived antibodies (VNARs) for highly specific hemoglobin detection. The T-12-4D VNAR shows exceptional affinity and stability, enabling accurate diagnostic assays for hemoglobin-related diseases.

Keywords:

Characterization Hemoglobin Phage display Screening VNAR

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Area of Science:

Biotechnology
Immunology
Molecular Biology

Background:

Hemoglobin is crucial for oxygen transport and diagnosing blood disorders.
Existing immunological detection methods require highly specific and high-affinity antibodies.
Shark-derived variable domain of new antigen receptor (VNAR) antibodies offer small size, stability, and high affinity for diagnostic applications.

Purpose of the Study:

To develop and characterize novel hemoglobin-specific VNAR antibodies.
To evaluate the affinity, specificity, and stability of these VNARs for potential diagnostic use.
To establish a diagnostic assay for human hemoglobin detection using the developed VNARs.

Main Methods:

Immunization of Chiloscyllium plagiosum with human hemoglobin and construction of VNAR immune libraries.
Phage display biopanning to identify hemoglobin-specific VNAR sequences (5-10C, 7-11A, T-12-4D).
Expression and purification of VNAR-Fc fusion proteins, followed by binding assays (ELISA, BLI), stability tests, and molecular dynamics simulations.

Main Results:

Three hemoglobin-specific VNAR sequences were identified.
T-12-4D-Fc demonstrated the highest affinity (K_D = 7.59 nM) and superior physicochemical stability under various stress conditions (heat, pH, urea).
A double-antibody sandwich ELISA (DAS-ELISA) using T-12-4D-Fc showed high accuracy and specificity for human hemoglobin detection in whole blood samples.

Conclusions:

The identified VNARs, especially T-12-4D, possess high affinity, specificity, and stability for hemoglobin detection.
These shark-derived antibodies fill a gap in targeting hemoglobin for diagnostic purposes.
The developed VNARs provide a foundation for improved diagnostics and monitoring of hemoglobin-related diseases.