Combining light- and heavy-chain-coding regions in a single bicistronic mRNA enables rapid isolation of mammalian cell clones expressing high levels of functional recombinant antibody.
May expedite high-yield recombinant antibody clone selection in research; leaves open translation to clinical manufacturing.
The variable regions of the murine monoclonal antibody A1, which effectively neutralizes the infection of susceptible cells by the murine hepatitis virus strain JHM, were cloned, sequenced, and expressed in mammalian cells as a functional recombinant antibody. To accomplish the concurrent synthesis of both antibody chains, the light- and heavy-chain-coding regions were inserted into a bicistronic expression cassette based on the encephalomyocarditis virus internal ribosomal entry site. The strategy of combining both coding regions in one bicistronic mRNA allows for the rapid isolation of cell clones expressing high levels of recombinant antibody.
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Kolb et al. (1997) studied this question.
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