ArticlemAbs
The influence of antibody fragment format on phage display based affinity maturation of IgG.
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Who cites it
62 citing papers in PubMed, 106 citations in OpenAlex.
- Integrating fab phage and yeast display through seamless transfer of paired heavy- and light-chain libraries for efficient antibody discovery.Antibody therapeutics · 2026Article
- Efficient generation of epitope-targeted antibodies with Germinal.Nature biotechnology · 2026Article
- Development of a Human IgG1 Monoclonal Antibody Targeting Transferrin Receptor 1 for Antitumor Drug Delivery.Antibodies (Basel, Switzerland) · 2026Article
- Phage Display Technology: Design, Library Construction, and Panning Strategies for Antibody Development.Biomolecules & therapeutics · 2026Review
- Applying recombinant antibodies for quality control of mixed-culture peanut bradyrhizobial inoculant.AMB Express · 2026Article
- Generation of a CDR H3 Randomized Monoclonal Antibody Library by Kunkel Mutagenesis.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Construction of Naïve and Immune Human Fab Phage Display Library.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Resolving the Interference of Anti-CD38 Antibodies on Blood Compatibility Assays Using CD38 "Baitbodies" Approach.Journal of immunology research · 2026Article
- Technological advancements in antibody-based therapeutics for treatment of diseases.Journal of biomedical science · 2025Review
- Targeting CD44 and EpCAM with Antibody Dye Conjugates for the Photoimmunotherapy of Prostate Cancer.Antibodies (Basel, Switzerland) · 2025Article
- Cancer Antibody Engineering: Comparison of Mammalian, Yeast, Bacterial, Plants, Cell-free and Hybridoma Expression Systems.Current pharmaceutical biotechnology · 2025Review
- Monoclonal antibodies: From magic bullet to precision weapon.Molecular biomedicine · 2024Review
- Human antibodies neutralizing the alpha-latrotoxin of the European black widow.Frontiers in immunology · 2024Article
- Genotyped functional screening of soluble Fab clones enables in-depth analysis of mutation effects.Scientific reports · 2023Article
- A mammalian cell display platform based on scFab transposition.Antibody therapeutics · 2023Article
- Microglia mediate neurocognitive deficits by eliminating C1q-tagged synapses in sepsis-associated encephalopathy.Science advances · 2023Article
- The variable conversion of neutralizing anti-SARS-CoV-2 single-chain antibodies to IgG provides insight into RBD epitope accessibility.Protein engineering, design & selection : PEDS · 2023Article
- Construction of Naïve and Immune Human Fab Phage Display Library.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Construction of Human Immune and Naive scFv Phage Display Libraries.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Antibody Phage Display.Methods in molecular biology (Clifton, N.J.) · 2023Article
2 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Today, most approved therapeutic antibodies are provided as immunoglobulin G (IgG), whereas small recombinant antibody formats are required for in vitro antibody generation and engineering during drug development. Particularly,single chain (sc) antibody fragments like scFv or scFab are well suited for phage display and bacterial expression, but some have been found to lose affinity during conversion into IgG.In this study, we compared the influence of the antibody format on affinity maturation of the CD30-specific scFv antibody fragment SH313-F9, with the overall objective being improvement of the IgG. The variable genes of SH313-F9 were randomly mutated and then cloned into libraries encoding different recombinant antibody formats, including scFv, Fab, scFabΔC, and FabΔC. All tested antibody formats except Fab allowed functional phage display of the parental antibody SH313-F9, and the corresponding mutated antibody gene libraries allowed isolation of candidates with enhanced CD30 binding. Moreover, scFv and scFabΔC antibody variants retained improved antigen binding after subcloning into the single gene encoded IgG-like formats scFv-Fc or scIgG, but lost affinity after conversion into IgGs.Only affinity maturation using the Fab-like FabΔC format, which does not contain the carboxy terminal cysteines, allowed successful selection of molecules with improved binding that was retained after conversion to IgG. Thus, affinity maturation of IgGs is dependent on the antibody format employed for selection and screening. In this study, only FabΔC resulted in the efficient selection of IgG candidates with higher affinity by combination of Fab-like conformation and improved phage display compared with Fab.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.