Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2003 Jul;10(4):680-5.
doi: 10.1128/cdli.10.4.680-685.2003.

Production and characterization of a human recombinant monoclonal Fab fragment specific for influenza A viruses

Alessandra Desogus  1 , Roberto Burioni, Angela Ingianni, Francesca Bugli, Raffaello Pompei, Giovanni Fadda
Affiliations

Production and characterization of a human recombinant monoclonal Fab fragment specific for influenza A viruses

Alessandra Desogus et al. Clin Diagn Lab Immunol. 2003 Jul.

Abstract

A human recombinant monoclonal Fab fragment that specifically recognizes all the influenza A virus strains tested was produced in transformed Escherichia coli using the phage display technique. No strain of influenza B virus reacted with it. It was purified after four cycles of panning and by a single passage through an immunoaffinity column. About 1 mg of pure monoclonal antibody was obtained from 1 liter of culture medium in 3 working days. The Fab fragment reacted with a viral 27-kDa protein, which could reasonably be a matrix protein. Indirect immunofluorescence tests performed on virus-infected MDCK cells showed that this Fab fragment was at least equally efficient as other commercial monoclonal antibody-based systems in detecting influenza A viral infections. The potential advantages of human recombinant Fabs on murine monoclonal antibodies are discussed.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
PAGE of purified Fab obtained from transformed E. coli clone 9. The clone Fab 9 appears as a clear band with a molecular weight (MW) of about 50,000 (lane 2); lane 1 contains size marker.
FIG. 2.
FIG. 2.
Western blot analysis of FLU-A virus PAGE protein profile using the Fab fragment from clone 9. Lane A, protein profile of FLU-A virus (strain WSN); lane B, Western blot using Fab clone 9; lane C, molecular marker. The arrow indicates a 27-kDa protein with affinity for the Fab fragment produced.
FIG. 3.
FIG. 3.
Amino acid sequence of both heavy and light chains of the anti-FLU-A virus Fab fragment produced in the recombinant E. coli clone 9.
FIG. 4.
FIG. 4.
Indirect immunofluorescence of FLU-A virus-infected MDCK cells using recombinant human Fab produced in E. coli clone 9 (A). (B and C) The same antibody was tested with FLU-B virus-infected cells (B) and in mock-infected cells (C).
See this image and copyright information in PMC

References

    1. Barbas III, C. F., A. S. Kang, R. A. Lerner, and S. J. Benkovic. 1991. Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc. Natl. Acad. Sci. USA 88:7978-7982. - PMC - PubMed
    1. Barbas III, C. F., J. E. Crowe, D. Cabala, T. M. Jones, S. L. Zbedee, B. R. Murphy, R. Chanock, and D. R. Burton. 1992. Human monoclonal Fab fragments derived from a combinatorial library bind to respiratory syncytial virus f glycoprotein and neutralize infectivity. Proc. Natl. Acad. Sci. USA 89:10164-10168. - PMC - PubMed
    1. Bethell, R., and P. Smith. 1998. Recent developments in sialidase inhibitors for the treatment of influenza. Drugs Future 23:1099-1109.
    1. Bucher, D. J., A. Mikhail, S. Popple, P. Graves, G. Meiklojohn, D. S. Hodes, K. Johansson, and P. E. Halonen. 1991. Rapid detection of type A influenza viruses with monoclonal antibodies to the M protein (M1) by enzyme-linked immunosorbent assay and time-resolved fluoroimmunoassay. J. Clin. Microbiol. 29:2484-2488. - PMC - PubMed
    1. Bugli, F., R. Bastidas, D. R. Burton, R. A. Williamson, M. Clementi, and R. Burioni. 2001. Molecular profile of a human monoclonal antibody Fab fragment specific for Epstein-Barr virus gp350/220 antigen. Hum. Immunol. 62:362-367. - PubMed

Publication types

MeSH terms

LinkOut - more resources