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After a 10 min stabilization time, the measurement was reset and the buffers were introduced sequentially from high to low pH and back to high pH to complete a full titration.
The calculated dilution factor was 5.25 and sequencing was performed as a full titration run without bead enrichment, i.e. the run was performed on a 40×75 plate, divided into 8 sectors (a titration run uses 4 of these sectors with different numbers of DNA molecules per bead i.e. 1,4,16, and 64 respectively).
If no complex formation was found, the peptide was assigned as a non-binder to the HLA molecule in question; conversely, if complex formation was found in the initial screening, a full titration of the peptide was performed to determine the affinity of binding.
In paired samples, a full titration of 4-fold dilutions of serum was used.
A full titration of Trip 3 and Trip 4 with pre-incubated TNR I10 was then performed (Fig. 3e).
The full titration curves showed that the serum IgA response to HA conjugated with CPE was consistently higher than to the HA alone.
Similar(43)
The values derived from full titrations and from experiments with a single concentration are in good agreement (Table 3).The latter procedure is to be preferred when fragment hits are ranked according to affinity after a screening campaign.
In other words, the fold-increases in Figure 3B were determined from a full stoichiometric titration where each titration was completely and fully saturated.
A larger number of titrations are shown here than in panel A to prevent panel A from being overcrowded; each fold-increase was determined by a full stoichiometric titration where each titration was completely and fully saturated.
The operational lifetime of an individual coated electrode was well in excess of that required to conduct a full herbicide titration (data not shown).
(B ) Example of a full pH titration series (spectra collected at pH 8.4 [blue], 7.5 [black], 7.0 [cyan], 6.7 [green], and 6.5 [red]).
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