Products/Services Used | Details | Operation |
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PCR Cloning and Subcloning> | C-terminal His-tagged Bi-ScFvs DNA sequences were synthesized (GenScript, Piscataway, NJ) and cloned into the pcDNA3.1(-) vector (Thermo Fisher Scientific) while the Bi-ScFv lacking the Cterminal His-tag was subcloned into an IgG1 Fc vector (InvivoGen) as well as IgG1 Fc vectors carrying knob-into-hole mutations | Get A Quote |
HIV-1 broadly neutralizing antibodies (bNAbs) are being explored as passively administered therapeutic and preventative agents. However, the extensively diversified HIV-1 envelope glycoproteins (Env) rapidly acquire mutations to evade individual bNAbs in monotherapy regimens. The use of a "single" agent to simultaneously target distinct Env epitopes is desirable to overcome viral diversity. Here, we report the use of tandem single-chain variable fragment (ScFv) domains of two bNAbs, specific for the CD4-binding site and V3 glycan patch, to form anti-HIV-1 bispecific ScFvs (Bi-ScFvs). The optimal Bi-ScFv crosslinks adjacent protomers within one HIV-1 Env spike and has greater neutralization breadth than ... More