%0 Journal Article
%A Urraca, Javier L.
%A Moreno-Bondi, María C.
%A Orellana, Guillermo
%A Sellergren, Börje
%A Hall, Andrew J.
%D 2007
%T Molecularly Imprinted Polymers as Antibody
Mimics in Automated On-Line Fluorescent
Competitive Assays
%U https://acs.figshare.com/articles/journal_contribution/Molecularly_Imprinted_Polymers_as_Antibody_Mimics_in_Automated_On_Line_Fluorescent_Competitive_Assays/2999257
%R 10.1021/ac070277i.s001
%2 https://acs.figshare.com/ndownloader/files/4700320
%K MIA
%K MIP
%K Pyrenemethylacetamido penicillanic acid
%K Competitive binding studies
%K penicillin G
%K antibiotic
%K penicillin G procaine salt
%K BLA
%K PAAP
%K PENG
%K polymer
%K Molecularly Imprinted Polymers
%K emission quantum yields
%K analysis
%X An automated molecularly imprinted sorbent based assay
(MIA) for the rapid and sensitive analysis of penicillin-type β-lactam antibiotics (BLAs) has been developed and
optimized. The polymers were prepared using penicillin
G procaine salt as template (PENGp) and a stoichiometric
quantity of a urea-based functional monomer to target the
single oxyanionic species in the template molecule. Highly
fluorescent competitors (emission quantum yields of 0.4−0.95), molecularly engineered to contain pyrene labels
while keeping intact the 6-aminopenicillanic acid moiety
for efficient recognition by the cross-linked polymers, have
been tested as analyte analogues in the competitive assay.
Pyrenemethylacetamido penicillanic acid (PAAP) was the
tagged antibiotic providing for the highest selectivity when
competing with PenG for the specific binding sites in the
molecularly imprinted polymer (MIP). Upon desorption
from the MIP, the emission signal generated by the PAAP
was related to the antibiotic concentration in the sample.
The 50% binding inhibition concentration of penicillin G
standard curves was at 1.81 × 10-6 M PENG, and the
detection limit was 1.97 × 10-7 M. The sensor showed a
dynamic range (normalized signal in the 20 to 80% range)
from 6.80 × 10-7 to 7.21 × 10-6 M (20−80% binding
inhibition) PENG in acetonitrile:HEPES buffer 0.1 M at
pH 7.5 (40:60, v/v) solutions. Competitive binding studies demonstrated various degrees of cross-reactivity with
penicillin-type β-lactam antibiotics such as ampicillin
(71%), oxacillin (66%), penicillin V (56%), amoxicillin
(13%), and nafcillin (46%) and a lower response to other
isoxazolyl penicillins such as cloxacillin (27%) and dicloxacillin (16%). The total analysis time was 14 min per
determination, and the MIP reactor could be reused for
more than 150 cycles without significant loss of recognition. The automatic MIA has been successfully applied
to the direct analysis of penicillin G in spiked urine
samples with excellent recoveries (mean value 92%).
Results displayed by comparative analysis of the optimized
MIA with a chromatographic procedure for penicillin G
showed excellent agreement between both methods.
%I ACS Publications