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<https://discovery.ucl.ac.uk/id/eprint/14474> <http://purl.org/dc/terms/title> "Protein and cell therapy for lecithin-cholesterol acyltransferase (LCAT) deficiency"^^<http://www.w3.org/2001/XMLSchema#string> .
<https://discovery.ucl.ac.uk/id/eprint/14474> <http://purl.org/ontology/bibo/abstract> "Lecithin-cholesterol acyltransferase (LCAT) is an enzyme principally secreted by the\r\nliver into the circulation where it esterifies cholesterol and plays a key role in high-\r\ndensity lipoprotein (HDL) metabolism. In familial and acquired (liver disease) LCAT\r\ndeficiency, the failure to esterify cholesterol causes many cellular and metabolic\r\ndisturbances. Here, I describe the purification of recombinant LCAT and assess two\r\napproaches to treat LCAT deficiency. Human LCAT cDNA was cloned into a\r\nselectable expression vector and used to generate a stably–transfected Chinese\r\nhamster ovary (CHO) cells secreting human LCAT tagged with 6 histidine residues.\r\nProductive clones were selected, monitoring LCAT activity by a modification of a\r\nradioactive enzymic assay for plasma, and the enzyme purified from culture medium\r\nby immobilised cobalt affinity chromatography. The pure LCAT, as judged by SDS-\r\nPAGE, was used to raise monoclonal antibodies in LCAT knockout mice for future\r\ndevelopment of a sensitive immunoassay. For therapy, I evaluated injection of pure\r\nLCAT into the peritoneal cavity of LCAT knockout mice using single and repeat\r\ndose regimes. LCAT activity was measurable in plasma post-injection and the\r\npercentage of esterified cholesterol increased, while agarose gel electrophoresis\r\nconfirmed a rise in HDL levels. In a second approach, I encapsulated the\r\nrecombinant CHO cells in biocompatible and semipermeable alginate-polylysine\r\nmicrocapsules using a syringe pump extrusion method. A study in vitro showed that,\r\nafter an initial lag phase, LCAT was secreted for over 90 days with the capsules\r\nremaining intact. These microencapsulated cells were implanted into peritoneal\r\ncavities of LCAT-deficient mice. LCAT activity was detected in mice plasma one\r\nweek post-implantation; the relative amount of esterified cholesterol was increased\r\nand lipoprotein profile was improved. I conclude that injection of recombinant\r\nenzyme or of encapsulated LCAT-secreting cells are feasible therapies for familial\r\nand acquired LCAT deficiency."^^<http://www.w3.org/2001/XMLSchema#string> .
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