Antimicrobial Peptides for Detection and Diagnostic Assays - Designing Receptors for the Next Generation of Biosensors - page 94

Fig. 3 Binding of Cy5-labeled LPS to cecropin A (1-8)-melittin (1-18) hybrid ( top panel ) and
indolicidin ( bottom panel ) peptides immobilized to four commercial microtiter plates
immobilization (e.g., amine-directed linkage to Immobilizer Amino ® and maleic
anhydride-activated microtiter plates) result in significant differences in the label-
ing efficiency of free primary amines and target capture. These results suggest that
the presentation and orientation of surface-bound AMPs are highly sensitive to the
choice of immobilization chemistry used to attach the peptides to microtiter plates.
Moreover, for surface-bound AMPs, peptide display is an insufficient predictor of
the effectiveness of target capture.
To meet the need for a high density, HTS AMP-based detection platform, we
developed a silane-based method for facile and controllable covalent immobiliza-
tion of AMPs to microtiter plates [ 52 ]. This approach allows for a wide variety of
peptide immobilization schemes to be applied to a single microtiter plate. Briefly,
inert polystyrene microtiter plates were modified with plasma treatment to generate
reactive hydroxyl moieties on the plate surface. The microtiter plates were then