Quantification of particle-conjugated or particle-encapsulated peptides on interfering reagent backgrounds
Particle-based technologies are increasingly being used in diagnostics and therapeutics. The particles employed in these applications are usually composed of polymers such as poly(lactide-co-glycolide) (PLG) and functionalized with peptides or proteins. Peptide or protein conjugation to particles is...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2014-07-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/000114190 |
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| author | Woon Teck Yap W. Kelsey Song Niharika Chauhan P. Nina Scalise Radhika Agarwal Stephen D. Miller Lonnie D. Shea |
| author_facet | Woon Teck Yap W. Kelsey Song Niharika Chauhan P. Nina Scalise Radhika Agarwal Stephen D. Miller Lonnie D. Shea |
| author_sort | Woon Teck Yap |
| collection | DOAJ |
| description | Particle-based technologies are increasingly being used in diagnostics and therapeutics. The particles employed in these applications are usually composed of polymers such as poly(lactide-co-glycolide) (PLG) and functionalized with peptides or proteins. Peptide or protein conjugation to particles is frequently achieved using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), while dimethyl sulfoxide (DMSO) is used to retrieve surface-attached or encapsulated peptides or proteins by solubilizing the particles. We examined strategies based on bicinchoninic acid (BSA), Coomassie Plus, and 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) assays for the quantification of surface-attached or encapsulated peptides or proteins. We determined that the CBQCA assay is a highly sensitive and accurate substitute for radioactivity-based assays that is suitable for measuring multiple particle-bound or particle-encapsulated peptides or proteins in the presence of EDC or PLG in DMSO, compounds that interfere with the more commonly used BSA and Coomassie Plus assays. Our strategy enables the accurate quantification of peptides or proteins loaded onto or into particles—an essential component of particle-based platform design for diagnostics and therapeutics. |
| format | Article |
| id | doaj-art-a53f1bba1663459495b31f42ab4dc47c |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2014-07-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-a53f1bba1663459495b31f42ab4dc47c2025-08-20T02:25:57ZengTaylor & Francis GroupBioTechniques0736-62051940-98182014-07-01571394410.2144/000114190Quantification of particle-conjugated or particle-encapsulated peptides on interfering reagent backgroundsWoon Teck Yap0W. Kelsey Song1Niharika Chauhan2P. Nina Scalise3Radhika Agarwal4Stephen D. Miller5Lonnie D. Shea61Department of Biomedical Engineering, Northwestern University, Evanston, IL2Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL1Department of Biomedical Engineering, Northwestern University, Evanston, IL2Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL2Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL5Departments of Microbiology-Immunology and Interdepartmental Immunobiology Center, Feinberg School of Medicine, Northwestern University, Evanston, IL2Department of Chemical and Biological Engineering, Northwestern University, Evanston, ILParticle-based technologies are increasingly being used in diagnostics and therapeutics. The particles employed in these applications are usually composed of polymers such as poly(lactide-co-glycolide) (PLG) and functionalized with peptides or proteins. Peptide or protein conjugation to particles is frequently achieved using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), while dimethyl sulfoxide (DMSO) is used to retrieve surface-attached or encapsulated peptides or proteins by solubilizing the particles. We examined strategies based on bicinchoninic acid (BSA), Coomassie Plus, and 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) assays for the quantification of surface-attached or encapsulated peptides or proteins. We determined that the CBQCA assay is a highly sensitive and accurate substitute for radioactivity-based assays that is suitable for measuring multiple particle-bound or particle-encapsulated peptides or proteins in the presence of EDC or PLG in DMSO, compounds that interfere with the more commonly used BSA and Coomassie Plus assays. Our strategy enables the accurate quantification of peptides or proteins loaded onto or into particles—an essential component of particle-based platform design for diagnostics and therapeutics.https://www.future-science.com/doi/10.2144/0001141903-(4-carboxybenzoyl)quinoline-2-carboxaldehydebicinchoninic acid1-ethyl-3-(3-dimethylaminopropyl)carbodiimidepoly(lactide-co-glycolide)nanoparticlesmicroparticles |
| spellingShingle | Woon Teck Yap W. Kelsey Song Niharika Chauhan P. Nina Scalise Radhika Agarwal Stephen D. Miller Lonnie D. Shea Quantification of particle-conjugated or particle-encapsulated peptides on interfering reagent backgrounds BioTechniques 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde bicinchoninic acid 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide poly(lactide-co-glycolide) nanoparticles microparticles |
| title | Quantification of particle-conjugated or particle-encapsulated peptides on interfering reagent backgrounds |
| title_full | Quantification of particle-conjugated or particle-encapsulated peptides on interfering reagent backgrounds |
| title_fullStr | Quantification of particle-conjugated or particle-encapsulated peptides on interfering reagent backgrounds |
| title_full_unstemmed | Quantification of particle-conjugated or particle-encapsulated peptides on interfering reagent backgrounds |
| title_short | Quantification of particle-conjugated or particle-encapsulated peptides on interfering reagent backgrounds |
| title_sort | quantification of particle conjugated or particle encapsulated peptides on interfering reagent backgrounds |
| topic | 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde bicinchoninic acid 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide poly(lactide-co-glycolide) nanoparticles microparticles |
| url | https://www.future-science.com/doi/10.2144/000114190 |
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