At 10 μM tafamidis in plasma, only the first binding site of each TTR tetramer is likely to be occupied.
Only CTL showing specificity for a single serotype or limited cross-reactivity (i.e. strong binding of the tetramer folded with the peptide of one serotype, weak binding of the other) remained.
Moreover, non-specific binding of the tetramer (as seen on the CD8-negative subset) also varied between the different laboratories.
We observed that a major naturally occurring escape variant with an amino acid substitution at position 5 (K5E) in the epitopic peptide substantially increased binding of the respective tetramer to CD14+ monocytes as compared to the tetramer refolded with the wild type tetramer (Figure 1).
However, this represents a simplified analysis of the data, because each phase presumably represents binding of multiple tetramers along the DNA.
Tetramer staining to dissect proper TCR formation was not possible with the 58 T-cell line due to unspecific binding of tetramer to the transferred CD8α molecule that was needed for the functional analysis of the TCRs.
Under some conditions (high SSB and/or low ionic strength), a second tetramer binds to each 70-base length, but at a rate 2 orders of magnitude slower than the rate of binding of the first tetramer.
By an ∼2:1 DCC-SSB Cy3-dT70 ratio, almost all the Cy3 was giving a peak at 7.4 S, consistent with tight binDCC-SSB Cy3-dT70nDCC-SSB Cy3-dT70ratioively slow exchalmost
Various methods have been used to identify activated T cells such as binding of MHC tetramers and expression of cell surface markers in addition to cytokine-based assays.
In tumor cells, deletions, mutations, methylations of p53 binding sites, or mutations of p53 such as p53mt135, disturb the binding of p53 tetramers to their binding sites, and this prevents the binding of the p300 complex to IGFBP-3 promoter.
