Table 2.

Summary of methods for kinetic measurements by biointeraction affinity chromatography

Method	Application conditions	Usable range of affinitiesa	Information obtainedb
Plate height measurements	Linear zonal elution	Weak to moderate affinity	kd
Peak profiling	Linear zonal elution	Weak to moderate affinity	kd,app or kd
Zonal elution peak fitting	Non-linear zonal elution	Weak to moderate affinity	kd,app or kd
Frontal analysis curve fitting	Non-linear frontal analysis	Moderate to strong affinity	ka,app or ka
Frontal analysis moment analysisc	Non-linear frontal analysisd	Moderate to strong affinity	ka,app or ka
Peak decay method	Non-linear zonal elution	Weak to moderate affinity	kd or k-1
Split peak method	Non-linear or linear zonal elutiond	Strong affinity	k1 or ka

^aWeak affinity, K_a < 10⁴ M^-1; moderate affinity, K_a = 10⁴ − 10⁶ M^-1; Strong affinity, K_a > 10⁶ M^-1.

^bSymbols and abbreviations: k_d, dissociation rate constant; k_d,app, apparent dissociation rate; k_a, association rate constant; k_a,app, apparent association rate constant; k_-1, reverse mass transfer rate constant for movement of a solute from the stagnant mobile phase to the flowing mobile phase in a column; k₁, forward mass transfer rate constant for movement of a solute from the flowing mobile phase to the stagnant mobile phase.

^cThis approach refers to the analysis of frontal analysis curves through the use of Eq. (10).

^dThe results in this method can be extrapolated to infinite dilution to correct for any concentration dependence in the results.