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On-line Continuous-flow Biochemical Interaction

Furthermore, when using 96 nM fluorescein-biotin and 32 nM streptavidin, an injection of 1 pmol of biotin results in an almost complete blocking of streptavidin and, consequently, the maximum peak height possible under the current conditions is about 95% of the highest point (3), indicating that apparent binding of biotin to streptavidin is in the order of 95%. [Pg.203]

Because the interaction between biotin and streptavidin is strong (fCa = 0.6 X 10 L mol ) with a relatively fast association rate (k+i = 2.4 x 10 L mol s ) and slow dissociation rate (k i = 0.4 x 10 s ), the reaction times are fast, i.e. 10-20 s. Furthermore, the addition of reporter ligand is performed only after the analyte protein reaction has taken place in coil I, avoiding a displacement reaction that would substantially increase the overall reaction time. [Pg.203]

Hence the reaction coil volumes were kept as small as possible to reduce bandbroadening, i.e. 17 pL and 33 pL, for coil I and coil II, respectively. [Pg.204]


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Biochemical interactions

Continuous flow

Continuous on-line

Flow lines

On-flow

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