title: Enrichment of Bruch's Membrane from Human Donor Eyes.
abstract: Age-related macular degeneration (AMD) is a leading cause of visual impairment in the developed world. The disease manifests itself by the destruction of the center of the retina, called the macula, resulting in the loss of central vision. Early AMD is characterised by the presence of small, yellowish lesions called soft drusen that can progress onto late AMD such as geographic atrophy (dry AMD) or neovascularisation (wet AMD). Although the clinical changes are well described, and the understanding of genetic influences on conferring AMD risk are getting ever more detailed, one area lacking major progress is an understanding of the biochemical consequences of genetic risk. This is partly due to difficulties in understanding the biochemistry of Bruch's membrane, a very thin extracellular matrix that acts as a biological filter of material from the blood supply and a scaffold on which the retinal pigment epithelial (RPE) cell monolayer resides. Drusen form within Bruch's membrane and their presence disrupts nutrient flow to the RPE cells. Only by investigating the protein composition of Bruch's membrane, and indeed how other proteins interact with it, can researchers hope to unravel the biochemical mechanisms underpinning drusen formation, development of AMD and subsequent vision loss. This paper details methodologies for enriching either whole Bruch's membrane, or just from the macula region, so that it can be used for downstream biochemical analysis, and provide examples of how this is already changing the understanding of Bruch's membrane biochemistry.
authors.affiliation: Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester; Centre for Advanced Discovery and Experimental Therapeutics, Central Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre.
authors.forename: Selina
authors.initials: S
authors.lastname: McHarg
authors.affiliation: Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester; Centre for Advanced Discovery and Experimental Therapeutics, Central Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre.
authors.forename: Nicole
authors.initials: N
authors.lastname: Brace
authors.affiliation: Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester; Centre for Advanced Discovery and Experimental Therapeutics, Central Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre.
authors.forename: Paul N
authors.initials: PN
authors.lastname: Bishop
authors.affiliation: Centre for Ophthalmology & Vision Sciences, Institute of Human Development, University of Manchester; Centre for Advanced Discovery and Experimental Therapeutics, Central Manchester University Hospitals NHS Foundation Trust, Manchester Academic Health Science Centre; simon.clark-3@manchester.ac.uk.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Clark