Molecular Characterization of Multivalent Bioconjugates by Size-Exclusion Chromatography with Multiangle Laser Light Scattering
| Autor: | Randolph S. Ashton, David V. Schaffer, Kevin E. Healy, Jacob Freas Pollock, Nikhil A. Rode |
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| Rok vydání: | 2012 |
| Předmět: |
Light
Molecular Sequence Data Size-exclusion chromatography Biomedical Engineering Multiangle light scattering Pharmaceutical Science Bioengineering engineering.material Article Ultraviolet visible spectroscopy Scattering Radiation Hedgehog Proteins Amino Acid Sequence Hyaluronic Acid Pharmacology chemistry.chemical_classification Bioconjugation Chromatography Organic Chemistry Polymer Combinatorial chemistry Refractometry Carbohydrate Sequence chemistry Chromatography Gel engineering Spectrophotometry Ultraviolet Biopolymer Peptides Biotechnology Macromolecule |
| Zdroj: | Bioconjugate Chemistry. 23:1794-1801 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/bc3000595 |
| Popis: | The degree of substitution and valency of bioconjugate reaction products are often poorly judged or require multiple time- and product-consuming chemical characterization methods. These aspects become critical when analyzing and optimizing the potency of costly polyvalent bioactive conjugates. In this study, size-exclusion chromatography with multiangle laser light scattering was paired with refractive index detection and ultraviolet spectroscopy (SEC-MALS-RI-UV) to characterize the reaction efficiency, degree of substitution, and valency of the products of conjugation of either peptides or proteins to a biopolymer scaffold, i.e., hyaluronic acid (HyA). Molecular characterization was more complete compared to estimates from a protein quantification assay, and exploitation of this method led to more accurate deduction of the molecular structures of polymer bioconjugates. Information obtained using this technique can improve macromolecular engineering design principles and help to better understand multivalent macromolecular interactions in biological systems. |
| Databáze: | OpenAIRE |
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