ArticleInternational journal of molecular sciences2020
Rapid and Effective Generation of Nanobody Based CARs using PCR and Gibson Assembly.
Article in International journal of molecular sciences, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 44 citations in OpenAlex.
- Antibody format matters: A comparative analysis of VHH and scFv domains reveals superior in vivo CAR T cell function with VHH domains.Molecular therapy : the journal of the American Society of Gene Therapy · 2026Article
- Cationic mRNA Lipid Nanoparticles for Ex Vivo NanoCAR-T Cell Engineering.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Nanobody-Engineered CLL-1 CAR T Cells: Optimizing Tumor-Specific Cytotoxicity and Minimizing Off-Tumor Toxicity.Cancer research communications · 2026Article
- Adoptive T Cell therapy for cancer and autoimmune disease: current advances, challenges, and future directions.Oncology reviews · 2026Article
- Advances in Nanotechnology for Lymphoma Treatment: Targeted Delivery, Immunomodulation, and TME-Responsive Therapy Strategies.International journal of nanomedicine · 2026Review
- Article
- Application of nanobody‑based CAR‑T in tumor immunotherapy (Review).International journal of molecular medicine · 2025Review
- Discovery and preclinical development of a SdAb-based CAR-T technology for targeting CD33 in AML.Molecular therapy. Oncology · 2025Article
- Nanobody-enhanced chimeric antigen receptor T-cell therapy: overcoming barriers in solid tumors with VHH and VNAR-based constructs.Biomarker research · 2025Review
- Discovery of nanobodies: a comprehensive review of their applications and potential over the past five years.Journal of nanobiotechnology · 2024Review
- CDR grafting and site-directed mutagenesis approach for the generation and affinity maturation of Anti-CD20 nanobody.Molecular biology reports · 2024Article
- The development of chimeric antigen receptor T-cells against CD70 for renal cell carcinoma treatment.Journal of translational medicine · 2024Article
- Development of nanobodies targeting hepatocellular carcinoma and application of nanobody-based CAR-T technology.Journal of translational medicine · 2024Article
- Nanobody-derived bispecific CAR-T cell therapy enhances the anti-tumor efficacy of T cell lymphoma treatment.Molecular therapy oncolytics · 2023Article
- Nanobodies: a new potential for prostate cancer treatment.Journal of cancer research and clinical oncology · 2023Review
- Article
- NANOBODIES®: A Review of Diagnostic and Therapeutic ApplicationsInternational journal of molecular sciences · 2023Review
- T-cells engineered with a novel VHH-based chimeric antigen receptor against CD19 exhibit comparable tumoricidal efficacy to their FMC63-based counterparts.Frontiers in immunology · 2023Article
- Nanobodies in cell-mediated immunotherapy: On the road to fight cancer.Frontiers in immunology · 2023Review
- BigDNA: Primer Design Software for Overlap-Based Assembly of Phage Genomes and Larger DNAs.PHAGE (New Rochelle, N.Y.) · 2022Article
Corrections and comments
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Authors and funding
16 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recent approval of chimeric antigen receptor (CAR) T cell therapy by the European Medicines Agency (EMA)/Federal and Drug Administration (FDA) and the remarkable results of CAR T clinical trials illustrate the curative potential of this therapy. While CARs against a multitude of different antigens are being developed and tested (pre)clinically, there is still a need for optimization. The use of single-chain variable fragments (scFvs) as targeting moieties hampers the quick generation of functional CARs and could potentially limit the efficacy. Instead, nanobodies may largely circumvent these difficulties. We used an available nanobody library generated after immunization of llamas against Cluster of Differentiation (CD) 20 through DNA vaccination or against the ectodomain of CD33 using soluble protein. The nanobody specific sequences were amplified by PCR and cloned by Gibson Assembly into a retroviral vector containing two different second-generation CAR constructs. After transduction in T cells, we observed high cell membrane nanoCAR expression in all cases. Following stimulation of nanoCAR-expressing T cells with antigen-positive cell lines, robust T cell activation, cytokine production and tumor cell lysis both in vitro and in vivo was observed. The use of nanobody technology in combination with PCR and Gibson Assembly allows for the rapid and effective generation of compact CARs.
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Registered trials
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.