ArticleBMC biotechnology2015
Generation and analysis of the improved human HAL9/10 antibody phage display libraries.
Article in BMC biotechnology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 61 papers.
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61 citing papers in PubMed, 130 citations in OpenAlex.
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- Integrated scFv identification and CAR T cell generation for AML targeting in vivo.International journal of cancer · 2026Article
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- Targeting BCL11B in CAR-engineered lymphoid progenitors drives NK-like cell development with prolonged anti-leukemic activity.Molecular therapy : the journal of the American Society of Gene Therapy · 2025Article
- Development of an optimized cell-based selection system for phage display libraries.Biology methods & protocols · 2025Article
- Combining Cellular Immunization and Phage Display Screening Results in Novel, FcγRI-Specific Antibodies.Viruses · 2024Article
- Construction of a Human Immune Library from Gallbladder Cancer Patients for the Single-Chain Fragment Variable (Antibodies (Basel, Switzerland) · 2024Article
- Human antibodies neutralizing the alpha-latrotoxin of the European black widow.Frontiers in immunology · 2024Article
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- Development of an inhibiting antibody against equine interleukin 5 to treat insect bite hypersensitivity of horses.Scientific reports · 2023Article
- Antibody Selection in Solution Using Magnetic Beads.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Antibodies to coagulase ofFrontiers in immunology · 2023Article
- Antibody Affinity and Stability Maturation by Error-Prone PCR.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Antibody Selection via Phage Display in Microtiter Plates.Methods in molecular biology (Clifton, N.J.) · 2023Review
- Construction of Human Immune and Naive scFv Phage Display Libraries.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Structure-guided and phage-assisted evolution of a therapeutic anti-EGFR antibody to reverse acquired resistance.Nature communications · 2022Article
- A Novel Synthetic Antibody Library with Complementarity-Determining Region Diversities Designed for an Improved Amplification Profile.International journal of molecular sciences · 2022Article
- Review
- Identification of New Antibodies Targeting Malignant Plasma Cells for Immunotherapy by Next-Generation Sequencing-Assisted Phage Display.Frontiers in immunology · 2022Article
1 more citing papers are in PubMed but not listed here.
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Authors and funding
12 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundAntibody phage display is a proven key technology that allows the generation of human antibodies for diagnostics and therapy. From naive antibody gene libraries - in theory - antibodies against any target can be selected. Here we describe the design, construction and characterization of an optimized antibody phage display library.
resultsThe naive antibody gene libraries HAL9 and HAL10, with a combined theoretical diversity of 1.5×10(10) independent clones, were constructed from 98 healthy donors using improved phage display vectors. In detail, most common phagemids employed for antibody phage display are using a combined His/Myc tag for detection and purification. We show that changing the tag order to Myc/His improved the production of soluble antibodies, but did not affect antibody phage display. For several published antibody libraries, the selected number of kappa scFvs were lower compared to lambda scFvs, probably due to a lower kappa scFv or Fab expression rate. Deletion of a phenylalanine at the end of the CL linker sequence in our new phagemid design increased scFv production rate and frequency of selected kappa antibodies significantly. The HAL libraries and 834 antibodies selected against 121 targets were analyzed regarding the used germline V-genes, used V-gene combinations and CDR-H3/-L3 length and composition. The amino acid diversity and distribution in the CDR-H3 of the initial library was retrieved in the CDR-H3 of selected antibodies showing that all CDR-H3 amino acids occurring in the human antibody repertoire can be functionally used and is not biased by E. coli expression or phage selection. Further, the data underline the importance of CDR length variations.
conclusionThe highly diverse universal antibody gene libraries HAL9/10 were constructed using an optimized scFv phagemid vector design. Analysis of selected antibodies revealed that the complete amino acid diversity in the CDR-H3 was also found in selected scFvs showing the functionality of the naive CDR-H3 diversity.
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