title: Ultrasensitive detection of endotoxins using computationally designed nanoMIPs.
abstract: Novel molecularly imprinted polymer nanoparticles (nanoMIPs) were designed for endotoxin from Escherichia coli 0111:B4, using computational modeling. The screening process based on binding energy between endotoxin and each monomer was performed with 21 commonly used monomers, resulting in the selection of itaconic acid, methacrylic acid and acrylamide as functional monomers due to their strong binding interaction with the endotoxin template. The nanoMIPs were successfully synthesized with functional groups on the outer surface to aid in the immobilization onto sensor surface. The solid phase photopolymerization approach used for the synthesis of nanoMIPs ranging from 200 to 235 nm in diameter. The limit of detection and KD were significantly improved when endotoxin samples were prepared using a novel triethylamine method. This improved the efficiency of gold nanoparticle functionalization by targeting the subunits of the endotoxin. Compared to the vancomycin MIP control, the endotoxin MIPs displayed outstanding affinity and selectivity towards the endotoxin with KD values in the range of 4.4-5.3 × 10(-10) M, with limits of detection of 0.44 ± 0.02 ng mL(-1) as determined by surface plasmon resonance (SPR) sensor when itaconic acid was used as the functional monomer. The MIP surface can be regenerated >30 times without significant loss of binding activity making this approach highly cost effective for expensive analyte templates. The combination of molecular modeling and solid phase synthesis enabled the successful synthesis of nanoMIPs capable of recognition and ultrasensitive detection of endotoxins using the highly sensitive SPR biosensor with triethylamine method.
authors.affiliation: Cranfield University, Cranfield, Bedfordshire MK43 0AL, England, UK; Technical University of Berlin, Straße des 17. Juni 124, Berlin 10623, Germany. Electronic address: zeynep.altintas@cranfield.ac.uk.
authors.forename: Zeynep
authors.initials: Z
authors.lastname: Altintas
authors.affiliation: Cranfield University, Cranfield, Bedfordshire MK43 0AL, England, UK.
authors.forename: Mohammed J
authors.initials: MJ
authors.lastname: Abdin
authors.affiliation: Cranfield University, Cranfield, Bedfordshire MK43 0AL, England, UK.
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Tothill
authors.affiliation: Cranfield University, Cranfield, Bedfordshire MK43 0AL, England, UK; University of Leicester, Leicester LE1 7RH, England, UK.
authors.forename: Kal
authors.initials: K
authors.lastname: Karim
authors.affiliation: Cranfield University, Cranfield, Bedfordshire MK43 0AL, England, UK. Electronic address: i.tothill@cranfield.ac.uk.
authors.forename: Ibtisam E
authors.initials: IE
authors.lastname: Tothill
mesh_terms: D000465:Algorithms; D004731:Endotoxins / Q000032:analysis*; D004926:Escherichia coli / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D054802:Molecular Imprinting; D015394:Molecular Structure; D053758:Nanoparticles / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*
chemical_list: D004731:Endotoxins; D011108:Polymers
keywords: Biosensor; Computational modeling; Endotoxins; Molecularly imprinting polymers (MIPs); Nanomaterials; Triethylamine