The concentration of brain homogenates with the Amicon Ultra Centrifugal filters

Accurately measuring the brain concentration of a neurotherapeutic is critical in determining its pharmacokinetic profile in vivo. Biologics are potential therapeutics for neurologic diseases and biologics fused to an antibody targeting a transcytosis receptor at the Blood-Brain Barrier, designated...

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Autores principales:	Joshua Yang, Rachita K. Sumbria
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Brain concentration Pharmacokinetics Blood-Brain Barrier (BBB) Enzyme-linked immunosorbent assay (ELISA) Science Q
Acceso en línea:	https://doaj.org/article/2b15fcb42d1443af8720d69815bb034a
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spelling	oai:doaj.org-article:2b15fcb42d1443af8720d69815bb034a2021-11-24T04:31:21ZThe concentration of brain homogenates with the Amicon Ultra Centrifugal filters2215-016110.1016/j.mex.2021.101584https://doaj.org/article/2b15fcb42d1443af8720d69815bb034a2021-01-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2215016121003745https://doaj.org/toc/2215-0161Accurately measuring the brain concentration of a neurotherapeutic is critical in determining its pharmacokinetic profile in vivo. Biologics are potential therapeutics for neurologic diseases and biologics fused to an antibody targeting a transcytosis receptor at the Blood-Brain Barrier, designated as antibody-biologic fusion proteins, are Blood-Brain Barrier penetrating neurotherapeutics. The use of sandwich immunosorbent assays to measure concentrations of antibody-biologic fusion proteins in brain homogenates has become increasingly popular. The raw brain homogenate contains many proteins and other macromolecules that can cause a matrix effect, potentially interfering with the limit of detection of such assays and reduce the overall sample signal. Further, the low sample loading volumes while running these assays can reduce the sample signal. Our aim was therefore to optimize the existing tissue sample preparation and processing to concentrate the sample to elevate the signal of the analyte. Here, we present a protocol for concentrating and increasing the signal of transferrin receptor antibody-biologic fusion proteins in mouse brain homogenates using the Amicon Ultra Centrifugal filters.• The presented method uses the Amicon Ultra Centrifugal filters to concentrate mouse brain tissue homogenates.• The concentrated brain tissue homogenates are then assayed using standard sandwich enzyme-linked immunosorbent assay (ELISA) protocols.• This method improves upon the traditional brain homogenization procedure and ELISA measurements for antibody-biologic fusion proteins by effectively concentrating brain tissue homogenates.Joshua YangRachita K. SumbriaElsevierarticleBrain concentrationPharmacokineticsBlood-Brain Barrier (BBB)Enzyme-linked immunosorbent assay (ELISA)ScienceQENMethodsX, Vol 8, Iss , Pp 101584- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Brain concentration Pharmacokinetics Blood-Brain Barrier (BBB) Enzyme-linked immunosorbent assay (ELISA) Science Q
spellingShingle	Brain concentration Pharmacokinetics Blood-Brain Barrier (BBB) Enzyme-linked immunosorbent assay (ELISA) Science Q Joshua Yang Rachita K. Sumbria The concentration of brain homogenates with the Amicon Ultra Centrifugal filters
description	Accurately measuring the brain concentration of a neurotherapeutic is critical in determining its pharmacokinetic profile in vivo. Biologics are potential therapeutics for neurologic diseases and biologics fused to an antibody targeting a transcytosis receptor at the Blood-Brain Barrier, designated as antibody-biologic fusion proteins, are Blood-Brain Barrier penetrating neurotherapeutics. The use of sandwich immunosorbent assays to measure concentrations of antibody-biologic fusion proteins in brain homogenates has become increasingly popular. The raw brain homogenate contains many proteins and other macromolecules that can cause a matrix effect, potentially interfering with the limit of detection of such assays and reduce the overall sample signal. Further, the low sample loading volumes while running these assays can reduce the sample signal. Our aim was therefore to optimize the existing tissue sample preparation and processing to concentrate the sample to elevate the signal of the analyte. Here, we present a protocol for concentrating and increasing the signal of transferrin receptor antibody-biologic fusion proteins in mouse brain homogenates using the Amicon Ultra Centrifugal filters.• The presented method uses the Amicon Ultra Centrifugal filters to concentrate mouse brain tissue homogenates.• The concentrated brain tissue homogenates are then assayed using standard sandwich enzyme-linked immunosorbent assay (ELISA) protocols.• This method improves upon the traditional brain homogenization procedure and ELISA measurements for antibody-biologic fusion proteins by effectively concentrating brain tissue homogenates.
format	article
author	Joshua Yang Rachita K. Sumbria
author_facet	Joshua Yang Rachita K. Sumbria
author_sort	Joshua Yang
title	The concentration of brain homogenates with the Amicon Ultra Centrifugal filters
title_short	The concentration of brain homogenates with the Amicon Ultra Centrifugal filters
title_full	The concentration of brain homogenates with the Amicon Ultra Centrifugal filters
title_fullStr	The concentration of brain homogenates with the Amicon Ultra Centrifugal filters
title_full_unstemmed	The concentration of brain homogenates with the Amicon Ultra Centrifugal filters
title_sort	concentration of brain homogenates with the amicon ultra centrifugal filters
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/2b15fcb42d1443af8720d69815bb034a
work_keys_str_mv	AT joshuayang theconcentrationofbrainhomogenateswiththeamiconultracentrifugalfilters AT rachitaksumbria theconcentrationofbrainhomogenateswiththeamiconultracentrifugalfilters AT joshuayang concentrationofbrainhomogenateswiththeamiconultracentrifugalfilters AT rachitaksumbria concentrationofbrainhomogenateswiththeamiconultracentrifugalfilters
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The concentration of brain homogenates with the Amicon Ultra Centrifugal filters

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