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Related Experiment Videos

Rabbit monoclonal Fab derived from a phage display library

M Foti1, F Granucci, P Ricciardi-Castagnoli

  • 1CNR Cellular and Molecular Pharmacology Center, Milano, Italy. mariaf@farma.csfic.mi.cnr.it

Journal of Immunological Methods
|August 6, 1998
PubMed
Summary

Researchers developed a novel method using phage display to create rabbit monoclonal antibodies (RmAb). This technique successfully produced a recombinant Fab fragment for use in direct enzyme immunoassays, offering a new approach for antibody production.

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Area of Science:

  • Immunotechnology
  • Molecular Biology
  • Biochemistry

Background:

  • Traditional methods for producing rabbit monoclonal antibodies (RmAb) via eukaryotic cell fusion are not routine.
  • Phage display technology offers an alternative for generating recombinant antibodies.

Purpose of the Study:

  • To apply phage display technology for producing a recombinant rabbit Fab molecule against the KLH antigen.
  • To develop a novel method for generating functional RmAb fragments.

Main Methods:

  • Utilized phage display to select rabbit Fab fragments against KLH.
  • Subcloned selected Fab fragments into an expression vector for fusion protein production.
  • Produced a fusion protein of rabbit Fab and bacterial alkaline phosphatase (phoA) in Escherichia coli.

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Main Results:

  • Successfully produced a recombinant rabbit Fab fragment targeting KLH.
  • Generated a fusion protein comprising rabbit Fab and phoA dimer.
  • Demonstrated the utility of crude conjugate extract in direct enzyme immunoassays for KLH.

Conclusions:

  • Phage display is a viable method for producing rabbit monoclonal antibody fragments.
  • The recombinant Fab-phoA fusion protein can be effectively used in direct enzyme immunoassays.
  • This approach provides an alternative to traditional RmAb production methods.