Using soybean agglutinin and agglutinin I (RCA120) as model analytes, the impact of polymer chain length and nanoparticle core size are evaluated, with chain length found to have a significant effect on signal generationhighlighting the need to control the macromolecular architecture to tune response. length found to have a significant effect on signal generationhighlighting the need Rabbit polyclonal to HHIPL2 to control the macromolecular architecture to tune response. With optimized systems, lectins are detectable at subnanomolar concentrations, comparable to antibody-based systems. Complete lateral (S)-(+)-Flurbiprofen flow devices are also assembled to show how these devices can be deployed in the real world. This work shows that glycan-binding can be a valuable tool in rapid diagnostics. agglutinin I]) was shown to be different. This provides the opportunity to introduce selectivity not just through the glycan, but also through macromolecular engineering, which is a unique feature ofthis technology. Guided by these results, complete diagnostic devices were fabricated and used to detect SBA in 10 min at concentrations as low as 5 g mL?1. 2.?Results and (S)-(+)-Flurbiprofen Discussion The primary aim of this work was to develop lateral flow technology to enable the sensitive detection of lectins, using glycosylated polymer-stabilized AuNPs, as an alternative to traditional antibody-based detection systems. To achieve this, an understanding of how particle/polymer structure impacts lateral flow performance was required. Therefore, a library-based screening approach was undertaken, with SBA chosen as the model lectin for detection. The precise chain length, surface glycan density, and particle size have been previously shown to be crucial in plasmonic (aggregation) glyco-assays, by modulating particle/analyte interactions and outcomes, while also ensuring colloidal stability in complex media.[42,43] Reversible additionCfragmentation chain (S)-(+)-Flurbiprofen transfer (RAFT) polymerization was used to synthesize a panel of poly(hydroxyethyl acrylamide)s (PHEA) using pentafluorophenyl-2-(dodecylthiocarbonothioylthio)-2-methylpropanoate (PFP-DMP) as the RAFT agent to install a pentafluorophenyl group at the x-ray photoelectron spectrum of 100% GalPHEA72 @AuNP16. E) Graphical representation of AuNP library illustrating the three variables of diameter, coating DP, and glycan density. Table 1 Polymers prepared for detecting SBA. (Figure 1D), and in the N 1scans (amine and amides have similar/overlapping binding energies so were not distinguishable), (S)-(+)-Flurbiprofen showing the presence of the PHEA, which were not present in the naked AuNP samples. Similarly, ether (XPS cannot easily distinguish ether from alcohol and are combined (S)-(+)-Flurbiprofen in the model employed here) peaks in the C 1scans were far larger in samples containing 100% sugar than in the citrate-stabilized AuNPs with no polymer functionalization. It is important to note the presence of carbonyls and carboxylic acid carbons are from atmospheric contaminants, and the presence of carbide likely from the silicon wafer particle interface. With this library of glycoparticles to hand, their function was screened in a lateral flow assay. Figure 2 shows the setup of the assay. A dipstick was made, where the test line (to capture the lectin analyte) was made by depositing 1 L of 1 1 mg mL?1 Gal1-3GalAgglutinin I (UEA, 0.05 mg mL?1, Figure 2E), a lectin with no affinity for GalNAc. Open in a separate window Figure 2 Schematic of dipstick lateral flow assay.A) Design of dipstick. B) Lateral flow with unfunctionalized BSA where particles flow without engaging the test line. C) Lateral flow with Gall-3GalAgglutinin I (RCA120), Agglutinin I, and WGA were purchased from Vector Laboratories. Gal1-3Gal em /em 1-4GlcNAc-BSA (3 atom spacer, NGP0334) was purchased from Dextra Laboratories. Ultrapure water used for buffers was MilliQ grade 18.2 m resistance. Representative Polymerization of 2-Hydroxyethyl Acrylamide PHEA40 as representative example. 2.0 g (17.37 mmol) of 2-hydroxyethyl acrylamide, 0.043 g (0.15 mmol) of ACVA, and 0.368 g (0.69 mmol) of PFP-DMP was added to 16 mL 1:1 toluene:methanol and degassed with nitrogen for 30 min. The reaction vessel was stirred and heated to 70 C for 2 h. The solvent was removed under vacuum. The crude product was dissolved in the minimum amount of methanol. Diethyl ether cooled in liquid nitrogen was added to the methanol to form a precipitate. The mixture was centrifuged for 2 min at 13.