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Updated: Jul 15, 2026
Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity
Published on: May 1, 2018
New Anti-RSV Nucleoprotein Monoclonal Antibody Pairs Discovered Using Rabbit Phage Display Technology
Pierre-Emmanuel Baurand1, Jérémy Balland1, Emilia Galli2
1Diaclone SAS-Part of Medix Biochemica Group, 6 Rue Dr Jean-François-Xavier Girod, BP 1985, 25000 Besançon, France.
Insights
Researchers developed new rabbit monoclonal antibodies (mAbs) for detecting human respiratory syncytial virus (hRSV). These antibodies show high potential for sensitive and rapid hRSV diagnosis, aiding early intervention in young children.
Area of Science:
- Immunology
- Virology
- Biotechnology
Background:
- Human respiratory syncytial virus (hRSV) is a significant cause of severe respiratory illness in infants and young children.
- Rapid and sensitive detection of hRSV is crucial for timely and effective clinical management.
- Current diagnostic methods may require improvement in speed and sensitivity for early intervention.
Purpose of the Study:
- To develop novel rabbit monoclonal antibodies (mAbs) targeting the nucleoprotein (NP) of hRSV.
- To evaluate the diagnostic potential of these newly developed antibodies for hRSV detection.
- To establish a foundation for advanced diagnostic tools, such as lateral flow assays.
Main Methods:
- Phage display technology was employed to generate a rabbit antibody fragment (Fab) library.
- Immunization of a rabbit with recombinant hRSV nucleoprotein (NP) initiated antibody development.
- Panning, ELISA screening, and reformatting to full IgG were performed to select and validate NP-specific antibodies.
- Pairing tests were conducted on recombinant and native hRSV samples.
Main Results:
- A rabbit Fab library with high insertion rate (>95%) and significant enrichment (100-fold) was successfully constructed.
- Twenty-eight NP-specific Fab candidates were identified through ELISA screening.
- Eleven pairs of reformatted full IgG antibodies demonstrated the ability to detect hRSV in native clinical lysates.
- The developed mAbs show high potential for sensitive hRSV detection.
Conclusions:
- Novel rabbit monoclonal antibodies targeting hRSV nucleoprotein have been successfully developed.
- These antibodies exhibit significant potential for sensitive and rapid detection of hRSV.
- Further validation using lateral flow testing with patient samples is recommended to advance diagnostic applications.
Abstract:
Human respiratory syncytial virus (hRSV) is one of the major contagious viruses and causes complicated respiratory issues, especially in young children. The sensitive and fast detection of hRSV is critical for taking the most effective actions. In the present study, rabbit antibodies against the hRSV nucleoprotein (NP) were developed using phage display technology. A female rabbit was immunized with an hRSV strain A2 recombinant NP. A Fab library was built and sorted during two successive panning rounds for strain B and the A2 NP (recombinant preparations), respectively. The choice of candidates was performed using ELISA on the two NP strains. The obtained library was 3 × 106 cfu/mL, with an insertion rate of >95%. The two panning rounds permitted an enrichment factor of 100. ELISA screening allowed us to obtain 28 NP-specific Fab candidates. Among them, 10 retained candidates were reformatted into rabbit full IgG; thereafter, pairing tests on the recombinant strains and native lysate samples were performed. After the pairing tests on the recombinant strains, 53 pairs were identified. Eleven pairs were identified as being able to detect RSVs from native lysates. This work presents new high-potential monoclonal antibodies mAbs (mAbs), which would benefit from lateral flow testing data with patient materials.
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