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Characterization of covalent crosslinking strategies for synthesizing DNA-based bioconjugates

Journal of biological engineering, 2019-07, Vol.13 (1), p.63-63, Article 63 [Peer Reviewed Journal]

COPYRIGHT 2019 BioMed Central Ltd. ;COPYRIGHT 2019 BioMed Central Ltd. ;The Author(s). 2019 ;ISSN: 1754-1611 ;EISSN: 1754-1611 ;DOI: 10.1186/s13036-019-0191-2 ;PMID: 31333759

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Title:
Characterization of covalent crosslinking strategies for synthesizing DNA-based bioconjugates
Author: Wickramathilaka, Malithi P ; Tao, Bernard Y
Subjects: Bioengineering ; Biomolecules ; Chromatography ; Composition ; Crosslinking ; DNA ; DNA synthesis ; EDC crosslinking ; Genetic research ; High performance liquid chromatography ; Ionization ; Liquid chromatography ; MALDI-TOF ; Methods ; Organic compounds ; Phosphates ; Polymer crosslinking ; Product yield ; Protein crosslinking ; Proteins ; Reaction mechanism ; RP-HPLC
Is Part Of: Journal of biological engineering, 2019-07, Vol.13 (1), p.63-63, Article 63
Description: An adapted strategy from the conventional 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) crosslinking method was developed to form a covalently coupled phosphoramidated single stranded DNA (ssDNA). Matrix assisted laser desorption ionization-time of flight (MALDI-TOF) results demonstrated that the phosphoramidated ssDNA conjugate is stable for several days, and that phosphoramidation occurred exclusively at the 5' phosphate of ssDNA. A reversed phase high-performance liquid chromatography (RP-HPLC) method with UV detection was developed to determine the yield of conjugates. The methods coefficients of variation (%CV) were less than 6%, and biases ranged from - 5.1 - 1.2%. The conjugate yield via the conventional EDC method was 68.3 ± 2.2%, while that of the adapted EDC/Imidazole method was 79.0 ± 2.4% ( = 10). This study demonstrates a convenient one pot strategy for crosslinking biological molecules.
Publisher: England: BioMed Central Ltd
Language: English
Identifier: ISSN: 1754-1611
EISSN: 1754-1611
DOI: 10.1186/s13036-019-0191-2
PMID: 31333759
Source: PubMed Central
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DOAJ Directory of Open Access Journals

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Characterization of covalent crosslinking strategies for synthesizing DNA-based bioconjugates

COPYRIGHT 2019 BioMed Central Ltd. ;COPYRIGHT 2019 BioMed Central Ltd. ;The Author(s). 2019 ;ISSN: 1754-1611 ;EISSN: 1754-1611 ;DOI: 10.1186/s13036-019-0191-2 ;PMID: 31333759

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