Evaluation of molecule-microbe interactions with capillary electrophoresis: procedures, utility and restrictions.
Understanding the interactions between molecules and living organisms is of paramount importance for the evaluation of pharmaceutical activity, chemical toxicity and all manner of microbiological studies. The capability of capillary electrophoresis (CE) in the evaluation of molecule-microbe interactions is examined in the present paper. The fundamental chemical concept of the binding or association constant for molecular systems measured in free solution is discussed for biological systems where microorganisms uptake or associate with molecules from their environment. The heterogeneity of the living organisms must be understood and accounted for including differences related to semantics such as concentration units and the nature of the associations between two entities and large differences in the size and number of microorganisms as compared to molecules. Finally, the added complexity and even inhomogeneity of a cell compared to most molecular systems must be considered and possibly controlled. The binding of specific molecules to viruses is discussed. CE can be utilized to quickly determine if a molecule binds very strongly or not at all to a cell (i.e., a binary yes/no answer). This could be useful for initial high-throughput screening purposes when using capillary arrays, for example. CE can be useful for determining unusual (large) molecule/microbe stoichiometries. Finally, CE can sometimes be used to determine the size of binding constants (K(RL)) within certain limits provided experimental conditions can be formulated that minimize problems of biological heterogeneity.
Berthod
Alain
A
Department of Chemistry, Iowa State University, Ames, IA 50011-3111, USA. berthod@univ-lyon1.fr
Rodriguez
Mike
M
Armstrong
Daniel W
DW
D018407:Bacterial Physiological Phenomena; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D008024:Ligands; D018406:Virus Physiological Phenomena
D008024:Ligands