A tool kit for rapid cloning and expression of recombinant antibodies.

ABSTRACT: 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/? and IgG1/?, as well as recombinant grass pollen allergen Phl p 7 specific fully human IgE/? and IgG4/? 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.

SUBMITTER: Dodev TS 

PROVIDER: S-EPMC4115235 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature