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Immunoassay for Quantitative Detection of Antibody Transcytosis Across the Blood-Brain Barrier In Vitro.
Sodja, Caroline; Callaghan, Deborah; Haqqani, Arsalan S; Stanimirovic, Danica B; Costain, Willard J; Jezierski, Anna.
Afiliación
  • Sodja C; National Research Council of Canada, Ottawa, ON, Canada.
  • Callaghan D; National Research Council of Canada, Ottawa, ON, Canada.
  • Haqqani AS; National Research Council of Canada, Ottawa, ON, Canada.
  • Stanimirovic DB; National Research Council of Canada, Ottawa, ON, Canada.
  • Costain WJ; National Research Council of Canada, Ottawa, ON, Canada.
  • Jezierski A; National Research Council of Canada, Ottawa, ON, Canada. anna.jezierski@nrc-cnrc.gc.ca.
Methods Mol Biol ; 2549: 345-357, 2022.
Article en En | MEDLINE | ID: mdl-35218529
Automated high-throughput immunoassays are emerging as reliable analytic techniques for the quantitative detection of proteins from a variety of sample types. Herein, we describe a method using the Protein Simple Wes capillary-based automated immunoassays platform for the quantification of His- and HA-tagged antibody transcytosis across an in vitro transwell blood-brain barrier (BBB) model. Compared to conventional ELISA, fluorescence, and Mass Spec-based detection approaches, Wes provides comparable datasets with additional information regarding size, aggregation, and potential degradation of samples before and after BBB transcytosis. In this chapter, we have benchmarked our Wes technique against ELISA and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), using known BBB crossing (FC5) and non-crossing (A20.1) single domain antibodies.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Barrera Hematoencefálica / Células Endoteliales Tipo de estudio: Diagnostic_studies Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2022 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Barrera Hematoencefálica / Células Endoteliales Tipo de estudio: Diagnostic_studies Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2022 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Estados Unidos