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Related Experiment Video

Updated: May 17, 2026

High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR

High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR

Published on: November 28, 2016

A rapid phage-free platform for antigen-specific VNAR screening with BATCH system.

Wenjie Tang^1,2, Jinming Hu¹, Yanping Zhu¹

¹MOE Key Laboratory of Rare Pediatric Diseases, Center for Medical Genetics, School of Life Sciences, Central South University, Changsha, Hunan, China.

Protein Science : a Publication of the Protein Society

|May 16, 2026

Summary

A novel phage-free bacterial two-hybrid (BATCH) platform enables rapid discovery of shark variable new antigen receptors (VNARs). This method accelerates the identification of stable VNARs for therapeutic and diagnostic applications.

Keywords:

Chiloscyllium plagiosum VNAR antigen‐specific single‐domain antibody bacterial two‐hybrid (BATCH)phage‐free screening

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Last Updated: May 17, 2026

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Published on: November 28, 2016

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Published on: April 27, 2011

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Detection of Neutralization-sensitive Epitopes in Antigens Displayed on Virus-Like Particle (VLP)-Based Vaccines Using a Capture Assay

Published on: February 10, 2022

Area of Science:

Biotechnology
Immunology
Molecular Biology

Background:

Shark-derived variable new antigen receptors (VNARs) are advantageous single-domain antibodies.
Conventional phage display methods for VNAR discovery can be complemented by alternative screening approaches.

Purpose of the Study:

To develop and validate a phage-free bacterial two-hybrid (BATCH) platform for rapid, antigen-specific VNAR discovery.
To streamline the identification of stable VNARs from immunized Chiloscyllium plagiosum.

Main Methods:

A bacterial two-hybrid (BATCH) system was employed using a cDNA library from immunized Chiloscyllium plagiosum spleen.
Screening involved dual lacZ and CmR readouts with enhanced reporter systems in BTH101 cells.
AlphaFold3 modeling and mutagenesis were used to analyze VNAR binding and function.

Main Results:

The BATCH platform identified two unique VNAR binders (VNAR1 and VNAR2) against Cre recombinase in 16 hours.
Mutagenesis studies confirmed the importance of the CDR3 loop and disulfide bond for VNAR affinity and function.
The platform demonstrated a 1- to 2-day screening phase for VNAR discovery.

Conclusions:

The phage-free BATCH platform offers a rapid and efficient alternative to conventional methods for VNAR discovery.
This system facilitates the identification of stable VNARs with potential therapeutic and diagnostic applications.
The study highlights the utility of simplified in vivo selection processes for antibody engineering.