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 Experiment Videos

Antibody humanization by framework shuffling.

William F Dall'Acqua1, Melissa M Damschroder, Jingli Zhang

  • 1Department of Antibody Discovery and Protein Engineering, MedImmune, Inc., One Medimmune Way, Gaithersburg, MD 20878, USA. dall'acquaw@medimmune.com

Methods (San Diego, Calif.)
|April 26, 2005
PubMed
Summary
This summary is machine-generated.

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

Safety and efficacy of decompressive craniectomy versus standard craniotomy for large acute epidural haematoma with tentorial herniation in China (PREDICT-AEDH): a nationwide, multicentre, open-label, parallel-group, randomised controlled trial.

The Lancet. Neurology·2026
Same author

Integrative profiling of glymphatic dysfunction in adolescent subthreshold depression.

Journal of affective disorders·2026
Same author

Case Report: Case of severe allergic reaction in a child caused by chlorhexidine skin disinfectant.

Frontiers in allergy·2026
Same author

Comparative Transcriptome and Sugar Metabolism Analysis Reveal Regulatory Networks During Bud Dormancy Release in <i>Prunus mume</i>.

Plants (Basel, Switzerland)·2026
Same author

Prognostic Significance of Inflammatory, Cellular, and Tumor-Specific Biomarkers in Patients With Pleural and Peritoneal Mesothelioma.

Clinical lung cancer·2026
Same author

Influence of metabolic syndrome on disease characteristics and activity in inflammatory bowel disease: A retrospective cohort study.

Medicine·2026

A novel framework shuffling technique rapidly humanized a mouse antibody targeting cancer-related EphA2. This method efficiently generated humanized antibodies retaining critical binding and functional activity for potential cancer therapies.

Area of Science:

  • Immunology
  • Biotechnology
  • Oncology

Background:

  • Monoclonal antibody B233 targets the human receptor tyrosine kinase EphA2, which is overexpressed in various cancer types.
  • EphA2 is a promising target for antibody-based cancer therapeutics.
  • Humanization of mouse antibodies is crucial to reduce immunogenicity for clinical applications.

Purpose of the Study:

  • To develop and validate a new antibody humanization technique called framework shuffling.
  • To generate humanized variants of the anti-EphA2 antibody B233.
  • To assess the binding affinity and functional activity of the humanized antibodies.

Main Methods:

  • Framework shuffling: fusing the six complementarity-determining regions (CDRs) of mAb B233 to diverse human germline frameworks.

Related Experiment Videos

  • Construction of combinatorial Fab libraries using shuffled frameworks.
  • Screening of libraries for antigen binding to EphA2.
  • Two-step selection process: sequential humanization of light and heavy chains.
  • Conversion of lead candidates to human IgG1 and characterization of binding and functional activity.

    • Main Results:

    • Several humanized anti-EphA2 antibody variants were identified that retained antigen binding.
    • Dissociation constants (Kd) for three fully humanized variants ranged from 3 to 48 nM.
    • The best humanized variant showed binding affinity within 5-fold of the parental mouse antibody.
    • Humanized antibodies demonstrated biochemical activity comparable to the parental antibody, including induction of EphA2 phosphorylation.

    Conclusions:

    • Framework shuffling is an effective and rapid method for antibody humanization without requiring rational design or structural information.
    • The technique successfully generated humanized antibodies with preserved binding affinity and functional activity against EphA2.
    • This approach facilitates the development of clinically viable antibody therapeutics targeting EphA2 for cancer treatment.