Biosensors fur use in detecting analytes, particularly small analytes such
as those having a molecular weight of less than 5,000 Daltons, are
disclosed which comprise a membrane and an electrode and a reservoir
defined therebetween, the membrane having an inner layer proximate the
electrode and an outer layer remote from the electrode comprising a
closely packed array of amphiphilic molecules, a plurality or ionophores,
and a plurality of membrane spanning lipids, the ionophores comprising
first and second half membrane spanning monomers, the first half membrane
spanning monomers being provided in the inner layer and being prevented
from lateral diffusion within the membrane and the second half membrane
spanning monomers being provided in the outer layer and being free to
diffuse laterally within the membrane, the second half membrane spanning
monomers having attached thereto a first receptor which is reactive with
the small analyte, wherein a carrier to which is attached a plurality of
the analyte is reversibly bound to the first receptor via the analyte. In
an alternative embodiment, the second half membrane spanning monomers are
attached through a carrier and/or linker group to at least one analyte,
and a receptor is reversibly bound to the second half membrane spanning
monomers via the analyte and said carrier and/or linker group.