Evaluation of Colorimetric BCA-Based Quantification of Hydrazide Groups on Magnetic Particles
Magnetic micro- and nanoparticles (MPs) are considered to provide excellent solid support for many immunoanalytical and bioaffinity applications, particularly when they contain hydrazide groups available for site-specific immobilization of various glycoproteins, such as immunoglobulin G and enzymes....
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| Format: | Article |
| Language: | English |
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Wiley
2018-01-01
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| Series: | Journal of Spectroscopy |
| Online Access: | http://dx.doi.org/10.1155/2018/5492893 |
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| author | Zuzana Svobodova Barbora Jankovicova Zdenek Plichta Daniel Horak Zuzana Bilkova |
| author_facet | Zuzana Svobodova Barbora Jankovicova Zdenek Plichta Daniel Horak Zuzana Bilkova |
| author_sort | Zuzana Svobodova |
| collection | DOAJ |
| description | Magnetic micro- and nanoparticles (MPs) are considered to provide excellent solid support for many immunoanalytical and bioaffinity applications, particularly when they contain hydrazide groups available for site-specific immobilization of various glycoproteins, such as immunoglobulin G and enzymes. To prepare a highly active bioaffinity carrier with sufficient binding capacity, knowledge as to the type and concentration of functional groups used for ligand binding is crucial. Described here is a simple, nontoxic method for rapid estimation of hydrazide functional groups bound to MPs using bicinchoninic acid (BCA). BCA kits are routinely used for colorimetric detection and quantification of total protein in liquid samples. In this study, the BCA reagent was applied for quantification of hydrazide groups on MPs. The approach was carried out using an adipic acid dihydrazide (ADH) solution and subsequently using various hydrazide-containing magnetic and nonmagnetic carriers differing in the density of hydrazide groups. The BCA test’s results obtained on the MPs were compared with those from conventional amino and hydrazide group quantification by the 2,4,6-trinitrobenzenesulfonic acid (TNBS) test. |
| format | Article |
| id | doaj-art-2bfc3591b8bd4b358a865708dd15b04f |
| institution | Kabale University |
| issn | 2314-4920 2314-4939 |
| language | English |
| publishDate | 2018-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Spectroscopy |
| spelling | doaj-art-2bfc3591b8bd4b358a865708dd15b04f2025-02-03T01:00:53ZengWileyJournal of Spectroscopy2314-49202314-49392018-01-01201810.1155/2018/54928935492893Evaluation of Colorimetric BCA-Based Quantification of Hydrazide Groups on Magnetic ParticlesZuzana Svobodova0Barbora Jankovicova1Zdenek Plichta2Daniel Horak3Zuzana Bilkova4Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, 532 10 Pardubice, Czech RepublicDepartment of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, 532 10 Pardubice, Czech RepublicInstitute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovsky Sq. 2, 162 06 Prague 6, Czech RepublicInstitute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovsky Sq. 2, 162 06 Prague 6, Czech RepublicDepartment of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Studentska 573, 532 10 Pardubice, Czech RepublicMagnetic micro- and nanoparticles (MPs) are considered to provide excellent solid support for many immunoanalytical and bioaffinity applications, particularly when they contain hydrazide groups available for site-specific immobilization of various glycoproteins, such as immunoglobulin G and enzymes. To prepare a highly active bioaffinity carrier with sufficient binding capacity, knowledge as to the type and concentration of functional groups used for ligand binding is crucial. Described here is a simple, nontoxic method for rapid estimation of hydrazide functional groups bound to MPs using bicinchoninic acid (BCA). BCA kits are routinely used for colorimetric detection and quantification of total protein in liquid samples. In this study, the BCA reagent was applied for quantification of hydrazide groups on MPs. The approach was carried out using an adipic acid dihydrazide (ADH) solution and subsequently using various hydrazide-containing magnetic and nonmagnetic carriers differing in the density of hydrazide groups. The BCA test’s results obtained on the MPs were compared with those from conventional amino and hydrazide group quantification by the 2,4,6-trinitrobenzenesulfonic acid (TNBS) test.http://dx.doi.org/10.1155/2018/5492893 |
| spellingShingle | Zuzana Svobodova Barbora Jankovicova Zdenek Plichta Daniel Horak Zuzana Bilkova Evaluation of Colorimetric BCA-Based Quantification of Hydrazide Groups on Magnetic Particles Journal of Spectroscopy |
| title | Evaluation of Colorimetric BCA-Based Quantification of Hydrazide Groups on Magnetic Particles |
| title_full | Evaluation of Colorimetric BCA-Based Quantification of Hydrazide Groups on Magnetic Particles |
| title_fullStr | Evaluation of Colorimetric BCA-Based Quantification of Hydrazide Groups on Magnetic Particles |
| title_full_unstemmed | Evaluation of Colorimetric BCA-Based Quantification of Hydrazide Groups on Magnetic Particles |
| title_short | Evaluation of Colorimetric BCA-Based Quantification of Hydrazide Groups on Magnetic Particles |
| title_sort | evaluation of colorimetric bca based quantification of hydrazide groups on magnetic particles |
| url | http://dx.doi.org/10.1155/2018/5492893 |
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