Validation of Selective Capture of Fimbriated Uropathogenic by a Label-free Engineering Detection System Using Mannosylated Surfaces.

Label-free detection of pathogens is of major concern to the microbiologist community. Most procedures require several steps and amplification techniques. Carbohydrates are well-established receptors for host-pathogen interactions, which can be amplified using glycodendritic architectures on the basis of multivalent binding interactions. Given that uropathogenic bacterial FimH is based on such mannopyranoside-binding interactions, we demonstrate herein that synthetic monomeric and trimeric thiolated α-d-mannosides can be effectively bound to gold substrate-functionalized self-assembled monolayers (SAMs) preactivated with maleimide functionalities. Mannosides grafted onto SAMs were followed using Quartz Crystal Microbalance with Dissipation (QCM-D). Binding recognition efficiency was first evaluated using the plant lectin from (ConA) also using QCM-D. We showed a direct correlation between the amount of mannoside bound and the lectin attachment. Even though there was less trimer bound (nM/cm) to the surface, we observed a 7-fold higher amount of lectin anchoring, thus further demonstrating the value of the multivalent interactions. We next examined the relative fimbriated selective adhesion/capture to either the monomeric or the trimeric mannoside bound to the surface. Our results established the successful engineering of the surfaces to show adhesion via specific mannopyranoside binding but unexpectedly, the monomeric derivative was more efficient than the trimeric analog, which could be explained by steric hindrance. This approach strongly suggests that it could be broadly applicable to other Gram-negative bacteria sharing analogous carbohydrate-dependent binding interactions.