Journal
SCIENTIFIC REPORTS
Volume 4, Issue -, Pages -Publisher
NATURE PORTFOLIO
DOI: 10.1038/srep05885
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Funding
- National Institute for Health Research (NIHR) Biomedical Research Centre based at Guy's and St Thomas' NHS Foundation Trust
- King's College London
- CR UK/EPSRC/MRC/NIHR KCL/UCL Comprehensive Cancer Imaging Centre [C1519/A10331]
- Cancer Research UK [C30122/A11527, C30122/A15774]
- KCL Experimental Cancer Medicine Centre
- National Institute for Health Research
- Welsh Assembly Government
- HSC R&D Office for Northern Ireland
- Chief Scientist Office, Scotland
- Overseas Research Students Award Scheme
- Medical Research Council [MR/L023091/1]
- Medical Research Council (UK)
- Asthma UK Medical Research Council
- Academy of Medical Sciences
- London Law Trust
- MRC [G1100090] Funding Source: UKRI
- Academy of Medical Sciences (AMS) [AMS-SGCL10-Josephs] Funding Source: researchfish
- Asthma UK [MRC-AsthmaUKCentre] Funding Source: researchfish
- Medical Research Council [G1000758B, G1100090, G1000758] Funding Source: researchfish
- National Institute for Health Research [CL-2012-17-005] Funding Source: researchfish
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Over the last four decades, molecular cloning has evolved tremendously. Efficient products allowing assembly of multiple DNA fragments have become available. However, cost-effective tools for engineering antibodies of different specificities, isotypes and species are still needed for many research and clinical applications in academia. Here, we report a method for one-step assembly of antibody heavy- and light-chain DNAs into a single mammalian expression vector, starting from DNAs encoding the desired variable and constant regions, which allows antibodies of different isotypes and specificity to be rapidly generated. As a proof of principle we have cloned, expressed and characterized functional recombinant tumor-associated antigen-specific chimeric IgE/kappa and IgG(1)/kappa, as well as recombinant grass pollen allergen Phl p 7 specific fully human IgE/lambda and IgG(4)/lambda antibodies. This method utilizing the antibody expression vectors, available at Addgene, has many applications, including the potential to support simultaneous processing of antibody panels, to facilitate mechanistic studies of antigen-antibody interactions and to conduct early evaluations of antibody functions.
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