Optimization of Microparticle Reagents to Collect and Detect Antibody

Bead reagents are used in a large number of assays in bioscience and biotechnology to collect and purify antibodies by immobilization. Bead-based immunoassays offer high-throughput analysis of multiple antibodies in a single sample. Although a variety of antibody-binding moieties on the collection b...

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Veröffentlicht in:Langmuir : the ACS journal of surfaces and colloids. - 1992. - 35(2019), 36 vom: 10. Sept., Seite 11717-11724
1. Verfasser: Ramirez, Katily (VerfasserIn)
Weitere Verfasser: Campbell, Elizabeth, Han, So-Yun, Buehler, Joseph, Phan, Thuong, Young Yoon, Hee, Lee, Ye Lim, Suresh, Tanvi, Sulchek, Todd
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2019
Zugriff auf das übergeordnete Werk:Langmuir : the ACS journal of surfaces and colloids
Schlagworte:Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Immunoglobulin G Indicators and Reagents Silicon Dioxide 7631-86-9
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520 |a Bead reagents are used in a large number of assays in bioscience and biotechnology to collect and purify antibodies by immobilization. Bead-based immunoassays offer high-throughput analysis of multiple antibodies in a single sample. Although a variety of antibody-binding moieties on the collection beads have been studied, the physical and material properties of collection beads have not been optimized to isolate specific antibodies over a broad range of concentrations from complex environments containing cells. We present a study of how to optimally use microparticles coated with protein G to collect low concentrations of IgG antibodies from complex solutions. We study the impact of bead material, bead size, incubation time, and protein G density to more efficiently collect antibodies and detect specific antibodies via fluorescent antigen labeling. The minimum detectable limit and the minimum incubation time for antibody collection are used as metrics to evaluate the collection parameters. We found that larger silica beads can capture more antibodies from a low concentration of sample, with a minimum incubation time of 60 min to equilibrium binding, resulting in a minimum detectable concentration of antibodies of 26 nM. We show that simple biophysical optimization of antibody collection reagents can be used to improve the collection of low concentrations of antibodies in complex environments. We demonstrate that the technology may be useful for monitoring antibody secretions from hybridoma cultures 
650 4 |a Journal Article 
650 4 |a Research Support, N.I.H., Extramural 
650 4 |a Research Support, U.S. Gov't, Non-P.H.S. 
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650 7 |a Indicators and Reagents  |2 NLM 
650 7 |a Silicon Dioxide  |2 NLM 
650 7 |a 7631-86-9  |2 NLM 
700 1 |a Campbell, Elizabeth  |e verfasserin  |4 aut 
700 1 |a Han, So-Yun  |e verfasserin  |4 aut 
700 1 |a Buehler, Joseph  |e verfasserin  |4 aut 
700 1 |a Phan, Thuong  |e verfasserin  |4 aut 
700 1 |a Young Yoon, Hee  |e verfasserin  |4 aut 
700 1 |a Lee, Ye Lim  |e verfasserin  |4 aut 
700 1 |a Suresh, Tanvi  |e verfasserin  |4 aut 
700 1 |a Sulchek, Todd  |e verfasserin  |4 aut 
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