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Updated: Aug 19, 2026
A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
Published on: February 16, 2014
Neutralizing human Fab fragments against measles virus recovered by phage display
Cristina de Carvalho Nicacio1, R Anthony Williamson, Paul W H I Parren
1Microbiology and Tumor Biology Center, Karolinska Institutet, S-171 77 Stockholm, Sweden.
Abstract:
Five human recombinant Fab fragments (Fabs) specific for measles virus (MV) proteins were isolated from three antibody phage display libraries generated from RNAs derived from bone marrow or splenic lymphocytes from three MV-immune individuals. All Fabs reacted in an enzyme-linked immunosorbent assay with MV antigens. In radioimmunoprecipitation assays two of the Fabs, MV12 and MT14, precipitated an approximately equal 80-kDa protein band corresponding to the hemagglutinin (H) protein from MV-infected Vero cell cultures, while two other Fabs, MT64 and GL29, precipitated an approximately equal 60-kDa protein corresponding the nucleocapsid (N) protein. In competition studies with MV fusion, H- and N protein-specific monoclonal antibodies (MAbs), the H-specific Fabs predominantly blocked the binding of H-specific MAbs, while the N-specific Fabs blocked MAbs to N. In addition, N-specific Fabs bound to denatured MV N protein in Western blotting. The specificity of the fifth Fab, MV4, could not be determined. By plaque reduction assays, three of the five Fabs, MV4, MV12, and MT14, exhibited neutralizing activity (80% cutoff) against MV (LEC-KI strain) at concentrations ranging between approximately 2 and 7 microg x ml(-1). Neutralization capacity against MV strains Edmonston and Schwarz was also detected, albeit at somewhat higher Fab concentrations. In conclusion, three neutralizing Fabs were isolated, two of them reactive against the H glycoprotein of MV and another reactive against an undefined epitope. This is the first study in which MV-neutralizing human recombinant Fab antibodies have been isolated from phage display libraries.
Insights
Researchers isolated three human recombinant antibody fragments (Fabs) that neutralize measles virus (MV). These Fabs target MV proteins, including the hemagglutinin (H) glycoprotein, offering potential for new antiviral therapies.
Area of Science:
- Immunology
- Virology
- Biotechnology
Background:
- Measles virus (MV) poses a significant public health threat.
- Development of novel antiviral agents is crucial for combating MV infections.
- Phage display technology offers a powerful platform for antibody discovery.
Purpose of the Study:
- To isolate and characterize human recombinant antibody fragments (Fabs) specific for measles virus (MV) proteins.
- To evaluate the neutralizing activity of these Fabs against MV.
- To establish a foundation for developing new MV-targeting therapeutics.
Main Methods:
- Generation of antibody phage display libraries from MV-immune individuals.
- Screening and isolation of MV-specific Fabs using enzyme-linked immunosorbent assay (ELISA), radioimmunoprecipitation assay (RIPA), and Western blotting.
- Assessment of neutralizing activity via plaque reduction assays against various MV strains.
Main Results:
- Five human recombinant Fabs specific for MV proteins were successfully isolated.
- Two Fabs (MV12, MT14) targeted the MV hemagglutinin (H) protein, and two (MT64, GL29) targeted the nucleocapsid (N) protein.
- Three Fabs (MV4, MV12, MT14) demonstrated significant neutralizing activity against MV strains, with two targeting the H glycoprotein.
Conclusions:
- This study reports the first isolation of MV-neutralizing human recombinant Fabs from phage display libraries.
- The identified Fabs, particularly those targeting the H protein, show promise for therapeutic applications against measles.
- The findings contribute to the development of novel strategies for controlling measles virus infections.
