The outer membrane of Gram-negative bacteria contains bacterial endotoxins known as Lipopolysaccharides (LPS). Owing to the strong immune responses induced in humans and animals, these large molecules have a strong toxic effect that can cause severe fever, hypotension, shock, and death. Endotoxins are often present in the environment and medical implants and represent undesirable contaminations of pharmaceutical preparations and medical devices. To overcome the limitations of the standard technique, novel methods for early and sensitive detection of LPS will be of crucial importance. In this work, an interesting approach for the sensitive detection of LPS has been realized by exploiting optical features of nanoplasmonic transducers supporting Localized Surface Plasmon Resonances (LSPRs). Ordered arrays of gold nano-prisms and nano-disks have been realized by nanospheres lithography. The realized transducers have been integrated into a simple and miniaturized lab-on-a-chip (LOC) platform and functionalized with specific antibodies as sensing elements for the detection of LPS. Interactions of specific antibodies anchored on protein A-modified sensor chips with the investigated analyte resulted in a spectral shift in the plasmonic resonance peak of the transducers. A good linear relationship between peak shifts and the LPS concentration has been demonstrated for the fabricated nano-structures with a detection limit down to 5 ng/mL. Integration with a proper microfluidic platform demonstrates the possibility of yielding a prototypal compact device to be used as an analytical test for quality determination of pharmaceutical products.
Nanoplasmonic biosensing approach for endotoxin detection in pharmaceutical field
Colombelli A.;Rizzato S.;Monteduro A. G.;Maruccio G.;
2021-01-01
Abstract
The outer membrane of Gram-negative bacteria contains bacterial endotoxins known as Lipopolysaccharides (LPS). Owing to the strong immune responses induced in humans and animals, these large molecules have a strong toxic effect that can cause severe fever, hypotension, shock, and death. Endotoxins are often present in the environment and medical implants and represent undesirable contaminations of pharmaceutical preparations and medical devices. To overcome the limitations of the standard technique, novel methods for early and sensitive detection of LPS will be of crucial importance. In this work, an interesting approach for the sensitive detection of LPS has been realized by exploiting optical features of nanoplasmonic transducers supporting Localized Surface Plasmon Resonances (LSPRs). Ordered arrays of gold nano-prisms and nano-disks have been realized by nanospheres lithography. The realized transducers have been integrated into a simple and miniaturized lab-on-a-chip (LOC) platform and functionalized with specific antibodies as sensing elements for the detection of LPS. Interactions of specific antibodies anchored on protein A-modified sensor chips with the investigated analyte resulted in a spectral shift in the plasmonic resonance peak of the transducers. A good linear relationship between peak shifts and the LPS concentration has been demonstrated for the fabricated nano-structures with a detection limit down to 5 ng/mL. Integration with a proper microfluidic platform demonstrates the possibility of yielding a prototypal compact device to be used as an analytical test for quality determination of pharmaceutical products.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
