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

Single chain Fab (scFab) fragment.

Michael Hust1, Thomas Jostock, Christian Menzel

  • 1Abteilung Biotechnologie, Institut für Biochemie und Biotechnologie, Technische Universität Braunschweig, Braunschweig, Germany. m.hust@tu-bs.de <m.hust@tu-bs.de>

BMC Biotechnology
|March 10, 2007
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

The landscape of antibody production systems: recombinant expression for research, diagnostics and therapy.

Frontiers in bioengineering and biotechnology·2026
Same author

German Society of Nuclear Medicine procedure guideline for brain perfusion SPECT using 99mTc-labelled radiopharmaceuticals (09/2025).

Nuklearmedizin. Nuclear medicine·2026
Same author

Overcoming lentiviral delivery limitations in hard-to-transduce suspension cells for genome-wide CRISPR screening.

Molecular therapy. Advances·2026
Same author

Phase I Study Evaluating a Monoclonal Fc-Silenced SARS-CoV-2 Antibody in Patients With Moderate-to-Severe COVID-19.

Clinical therapeutics·2026
Same author

Resolving the Interference of Anti-CD38 Antibodies on Blood Compatibility Assays Using CD38 "Baitbodies" Approach.

Journal of immunology research·2026
Same author

Immunogenic <i>Streptococcus equi</i> cell surface proteins identified by ORFeome phage display.

mSphere·2025
Same journal

Editorial Expression of Concern: Hindering the biofilm of microbial pathogens and cancer cell lines development using silver nanoparticles synthesized by epidermal mucus proteins from Clarias gariepinus.

BMC biotechnology·2026
Same journal

Computer-guided enzyme engineering of PET hydrolase mutants towards improved PET affinity.

BMC biotechnology·2026
Same journal

Synthesis and application of exopolysaccharide-selenium nanocomposite produced by Pseudoduganella armeniaca ZMN-3.

BMC biotechnology·2026
Same journal

Read-mapping signatures for molecular characterization of transgenic DNA insertions in paired-end Illumina data.

BMC biotechnology·2026
Same journal

Comparative analysis of MinION and MiSeq using 16S rRNA gene amplicon sequencing in human gut microbiome.

BMC biotechnology·2026
Same journal

Integrated structure-based and systems-level identification of PDE9A inhibitors for heart failure with preserved ejection fraction.

BMC biotechnology·2026
See all related articles

A novel single-chain Fab fragment (scFab) enhances functional antibody fragment production in bacteria. This improved antibody design is suitable for both soluble expression and antibody phage display, advancing proteome binder generation.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Immunology

Background:

  • Single-chain variable fragments (scFv) enabled functional antibody fragment production in E. coli.
  • Fragment antigen-binding (Fab) fragments yield lower in E. coli due to size and assembly complexity.
  • A need exists for antibody designs balancing Fab stability with scFv expression levels for E. coli and phage display.

Purpose of the Study:

  • To develop a novel antibody format for improved production of functional antibody fragments.
  • To combine the stability of Fab fragments with high-level bacterial expression.
  • To create an antibody fragment suitable for soluble expression and antibody phage display.

Main Methods:

  • Introduced a polypeptide linker between the fragment difficult (Fd) and light chain (LC) to create single-chain Fab fragments (scFab).

Related Experiment Videos

  • Tested various linker designs and modifications of constant regions for phage display and soluble expression.
  • Expressed scFab variants in E. coli and Pichia pastoris, characterizing monovalent, divalent (DiFabodies), and multimeric forms.

    • Main Results:

    • A scFab variant lacking cysteines (scFabDeltaC) showed optimal phage display efficiency and soluble production.
    • The scFab format demonstrated successful expression in both E. coli and Pichia pastoris.
    • Characterized monovalent, divalent (DiFabodies), and multimeric antibody fragments.

    Conclusions:

    • A new antibody design, scFab, facilitates the generation of bivalent Fab derivatives.
    • This format is valuable for antibody phage display and soluble antibody fragment production.
    • The scFab format is particularly useful for high-throughput proteome binder generation due to avidity and standard detection methods.