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Multiplex bead binding assays using off-the-shelf components and common flow cytometers.
Hattori, Takamitsu; Koide, Akiko; Panchenko, Tatyana; Romero, Larizbeth A; Teng, Kai Wen; Corrado, Alexis D; Koide, Shohei.
  • Hattori T; Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016, USA; Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA.
  • Koide A; Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016, USA; Department of Medicine, New York University School of Medicine, New York, NY 10016, USA.
  • Panchenko T; Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016, USA.
  • Romero LA; Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA.
  • Teng KW; Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016, USA.
  • Corrado AD; Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016, USA.
  • Koide S; Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016, USA; Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA. Electronic address: Shohei.Koide@nyulangone.org.
J Immunol Methods ; 490: 112952, 2021 03.
Article in English | MEDLINE | ID: covidwho-1065340
ABSTRACT
The ability to quantify protein-ligand interactions in an accurate and high-throughput manner is important in diverse areas of biology and medicine. Multiplex bead binding assays (MBBAs) are powerful methods that allow for simultaneous analysis of many protein-ligand interactions. Although there are a number of well-established MBBA platforms, there are few platforms suitable for research and development that offer rapid experimentation at low costs and without the need for specialized reagents or instruments dedicated for MBBA. Here, we describe a MBBA method that uses low-cost reagents and standard cytometers. The key innovation is the use of the essentially irreversible biotin-streptavidin interaction. We prepared a biotin-conjugated fluorescent dye and used it to produce streptavidin-coated magnetic beads that are labeled at distinct levels of fluorescence. We show the utility of our method in characterization of phage-displayed antibodies against multiple antigens of SARS-CoV-2, which substantially improves the throughput and dramatically reduces antigen consumption compared with conventional phage ELISA methods. This approach will make MBBAs more broadly accessible.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Spike Glycoprotein, Coronavirus / COVID-19 Serological Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: J Immunol Methods Year: 2021 Document Type: Article Affiliation country: J.jim.2020.112952

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Spike Glycoprotein, Coronavirus / COVID-19 Serological Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: J Immunol Methods Year: 2021 Document Type: Article Affiliation country: J.jim.2020.112952