Singlechain antibodies (scA) were originally developed for Escherichia coli expression to bypass the costly production of monoclonal antibodies in tissue culture or mice.7273 They comprise only the variable domains of both the heavy chain and the light chain of an antibody. These domains are expressed from a single gene, in which the coding regions for these domains are separated by a short spacer sequence coding for a peptide bridge which connects the two variabledomain peptides. The resulting singlechain antibody (scA, also termed singlechain variable fragment, scFv) can bind to its antigen with similar affinity as an Fab fragment of the authentic antibody molecule.
scFvs have been developed by our group and others to combat HIV-1 replication when expressed intracellularly.237074-80 Both preintegration (e.g., integrase, reverse transcriptase, matrix protein) and postintegration (e.g., Rev and Tat) sites of the viral life cycle have been targeted, with varying success. Further studies utilizing constructs combining multiple scFvs for potential synergistic antiviral potency are under analysis and should be important in developing robust anti-HIV-1 molecular therapeutics.
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