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The increased binding valency of these scFv multimers results in high avidity (low off-rates).
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To further examine the role of CD3 binding valency in a non-specific or 'conditional' T cell activation, we examined the ability of DVD-Ig or Half DVD-Ig to stimulate a NFAT reporter in Jurkat T cells in the presence or absence of tumor cell associated antigen.
To understand if we are able to generate Half DVD-Ig molecules to study the effect of anti-CD3 binding valency on redirected T cell cytotoxicity for tumor killing in vitro, we generated a Half anti-EGFR (outer domain)/anti-CD3 (inner domain) DVD-Ig molecule connected using a long-long (L-L) linker by including four mutations at CH3 domain and two cysteine mutations at the hinge region (Fig. 2A).
Binding strength or avidity of macromolecules is governed by three major factors: the intrinsic affinity of the individual binding interface, the valency of the binding site(s), and geometric arrangement of the interacting components [11], [18].
Transcription factor binding site enrichment analysis verified the binding of these MRs to their predicted targets.
The stability of the complex formed by this bispecific targeting adapter and the Ad virion resulted in insufficient gene transfer and was subsequently improved by increasing the valency of adapter virus binding.
The precise sequences allowed the group to assess specifically how the valency of the binding motif influenced the strength of cell adhesion under shear stress.
This approximately tenfold difference enables detection of lower concentrations of the binding partner PD-1, possibly due to the higher flexibility and valency of the IgM backbone promoting binding and detection.
A possible mechanism that could facilitate binding or reduce dissociation might be the higher valency of the IgM-backbone.
Soon after the description of the first TNFRSF receptor-specific agonistic antibodies, it turned out that the valency of antibodies, thus the antigen binding sites per molecule, is of crucial relevance for the agonistic activity.
In order to guide experimental design, a kinetic model was built to explore how the affinity and valency of dual-ligand liposomes affect the binding and selectivity of delivery to cells with various receptor expression.
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