Advanced methodology for LPS capture from biofluids
ABSTRACT: Lipopolysaccharide (LPS), or endotoxin, is the main component of the outer membrane of Gram-negative bacteria where lipid A is the responsible segment for its toxicity. It poses a serious risk when detected both in different industries and environments and when is present in human bloodstr...
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| Tipo de recurso: | tesis doctoral |
| Fecha de publicación: | 2021 |
| País: | España |
| Institución: | Universidad de Cantabria (UC) |
| Repositorio: | UCrea Repositorio Abierto de la Universidad de Cantabria |
| Idioma: | inglés |
| OAI Identifier: | oai:repositorio.unican.es:10902/21926 |
| Acceso en línea: | http://hdl.handle.net/10902/21926 |
| Access Level: | acceso abierto |
| Palabra clave: | Toxin sequestration Lipopolysaccharides (LPS) Protein design and engineering Microfluidics Computational fluid dynamics (CFD) Captura de toxinas Lipopolisacáridos (LPS) Diseño e ingeniería de proteínas Microfluidica Dinámica computacional de fluidos (CFD) |
| Sumario: | ABSTRACT: Lipopolysaccharide (LPS), or endotoxin, is the main component of the outer membrane of Gram-negative bacteria where lipid A is the responsible segment for its toxicity. It poses a serious risk when detected both in different industries and environments and when is present in human bloodstream as it can lead to sepsis, an exaggerated response to LPS that triggers immune suppression, organ dysfunction or even death. Unfortunately, alternative methods for contaminant removal through extracorporeal blood detoxification processes present drawbacks that make endotoxin detection/removal a crucial challenge to achieve safe and effective detoxification processes. In this regard, magnetofluidic devices deserve special attention and involve two main steps: the sequestration of LPS on suitably functionalized magnetic nanoparticles (MNPs) and, the removal of the MNPs-LPS complex from the biological fluid. Consequently, this dissertation provides an integrated methodology to advance the design of the LPS sequestration step to promote its separation from biofluids through the synthesis of an antilipopolysaccharide (LALF) protein from Limulus polyphemus species using genetic engineering techniques in addition to the quantification of the binding strength of the LALF protein to LPS through a newly approach and addressing the variables that affect the formation of the complex. In addition, in order to contribute to the development of an application for continuous LPS capture, as a first approach, homogeneous and heterogeneous L-L separation of aqueous anions (chromate) in microdevices is addressed experimentally and by means of a theoretical model developed with ANSYS FLUENT, laying the foundations to continue with the microfluidic design for L-S separation and finally, its application to LPS capture. |
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