Kinetic modeling and analysis of a vesicle system for immunosensor development

  • Yong-Yi Y
  • Van Wie B
  • Koch A
 et al. 
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A novel mechanism is presented for immunosensor development that uses an immunological competition reaction in a vesicle system. This system consists of a suspension of reconstituted vesicles, channel agonist, protein linker to block the channels, voltage sensitive dye and analyte to be detected. In the proposed mechanism analyte serves a catalytic role as individual analytes competitively displace multiple channel linkers through association with one channel, dissociation and new associations with other channels. When one channel opens on a vesicle a permanent Nernst potential develops for that vesicle leading to fluorescence of voltage sensitive dyes. The time constant of the redistribution from linker-channel form to analyte-channel form is 0.92/k4(k4is the off-rate constant for the analyte-channel association) in the region of analyte concentrations less than 10-9M. Kinetic analyses show that several factors, including concentration of analyte or linker, number of channels per vesicle, on-rate or off-rate constant of the linker-channel and on-rate constant of analyte-channel complexes have significant effects on the minimum signal response time.

Author-supplied keywords

  • Antibody-antigen binding kinetics
  • Immunosensors
  • Off-rate constants
  • On-rate constants
  • Protein linker
  • Receptor channels
  • Reconstituted vesicles

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