Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Targeting mGlyR with nanobodies for depression.

Nature communications·2026
Same author

Engineered antibodies that stabilize drug-modified KRAS<sup>G12C</sup> neoantigens enable selective and potent cross-HLA immunotherapy.

Nature communications·2025
Same author

Potentiating CD20 monoclonal antibody therapy by targeting complement C3 fragments covalently deposited on lymphoma cells.

Blood·2025
Same author

Generation of Antibody Libraries for Phage Display: Preparation of Electrocompetent <i>E. coli</i>.

Cold Spring Harbor protocols·2024
Same author

Phage Display Selection of Antibody Libraries: Screening of Selected Binders.

Cold Spring Harbor protocols·2024
Same author

Generation and Selection of Phage Display Antibody Libraries in Fab Format.

Cold Spring Harbor protocols·2024

Related Experiment Video

Updated: Jun 10, 2025

Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
12:55

Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries

Published on: January 17, 2015

18.5K

Generation of Antibody Libraries for Phage Display: Library Reamplification.

Haiyong Peng1, Christoph Rader2

  • 1Department of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, Jupiter, Florida 33458, USA.

Cold Spring Harbor Protocols
|October 15, 2024
PubMed
Summary

Reamplifying phage display libraries (Fab libraries) is crucial for antibody discovery. This protocol details reamplification steps, ensuring library diversity is maintained for effective antibody selection and evolution.

More Related Videos

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
10:17

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Published on: January 14, 2020

7.7K
Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity
12:31

Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity

Published on: May 1, 2018

13.6K

Related Experiment Videos

Last Updated: Jun 10, 2025

Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
12:55

Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries

Published on: January 17, 2015

18.5K
Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
10:17

Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library

Published on: January 14, 2020

7.7K
Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity
12:31

Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity

Published on: May 1, 2018

13.6K

Area of Science:

  • Biotechnology
  • Immunology
  • Molecular Biology

Background:

  • Phage display technology is vital for antibody discovery and engineering.
  • Fab libraries, containing diverse antibody fragments, are used for selecting specific binders.
  • Maintaining library diversity during antibody selection is critical for success.

Purpose of the Study:

  • To describe a protocol for reamplifying original phage display Fab libraries.
  • To address the practical necessity of reamplification despite potential biases.
  • To provide guidelines for reamplifying libraries from early and later selection rounds.

Main Methods:

  • Detailed step-by-step protocol for reamplifying Fab-phage libraries.
  • Discussion on the impact of reamplification on library diversity.
  • Considerations for using freshly prepared vs. reamplified Fab-phage.

Main Results:

  • The described protocol enables the reamplification of Fab-phage libraries.
  • Reamplification can introduce bias towards higher-expressing clones.
  • The protocol is applicable to original libraries and libraries from later panning rounds.

Conclusions:

  • Reamplification of Fab-phage libraries is often a necessary step in antibody discovery workflows.
  • The presented protocol provides a method to reamplify libraries while acknowledging potential biases.
  • Careful consideration of reamplification is needed to preserve library integrity for antibody evolution.