Details
| Original language | English |
|---|---|
| Pages (from-to) | 65-69 |
| Number of pages | 5 |
| Journal | Cytometry Part A |
| Volume | 62A |
| Issue number | 1 |
| Publication status | Published - 11 Oct 2004 |
Abstract
Background: A new method was established to characterize the binding kinetics of DNA toward layered double hydroxides (LDHs). The setup consisted of a newly developed sampling tube that allows the injection of analyte during the flow cytometric measurement. Methods: Layered double hydroxides consist of cationic metal hydroxide layers and exchangeable interlayer anions. This negatively charged structure permits biomolecules such as DNA to adsorb, and a so-called DNA-LDH hybrid is formed. The hydroxide layers can be removed in acidic media and the DNA will be released. CERATOFIX® (a registered trademark of SüdChemie AG NA that belongs to the family of LDHs, produced by Süd-Chemie AG). The chemical structure can be summarized as [Mg 2Al(OH)6](CO3)0.5. The binding capacity and kinetic characteristics of different types of CERATOFIX®NA for a model DNA was determined by flow cytometry. Results: The static binding capacities of the different LDHs were determined after 1- and 16-h incubation with DNA solution, showing different binding patterns between the LDH materials. The binding kinetics were revealed by flow cytometric measurements in short-term and long-term kinetic experiments, showing that the majority of DNA adsorbs within the first 60 s. Conclusions: DNA removal from cell culture supernatants is one of the major concerns in downstream processing. Due to the anion exchange capabilities of LDHs it seemed a very interesting approach to use these materials for binding of DNA for elimination purposes.
Keywords
- DNA removal, Flow cytometry, LDH
ASJC Scopus subject areas
- Medicine(all)
- Pathology and Forensic Medicine
- Medicine(all)
- Histology
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: Cytometry Part A, Vol. 62A, No. 1, 11.10.2004, p. 65-69.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Flow Cytometry: Interesting Tool for Studying Binding Behavior of DNA on Inorganic Layered Double Hydroxide (LDH)
AU - Burzlaff, Arne
AU - Brethauer, Simone
AU - Kasper, Cornelia
AU - Jackisch, Björn Oliver
AU - Scheper, Thomas
PY - 2004/10/11
Y1 - 2004/10/11
N2 - Background: A new method was established to characterize the binding kinetics of DNA toward layered double hydroxides (LDHs). The setup consisted of a newly developed sampling tube that allows the injection of analyte during the flow cytometric measurement. Methods: Layered double hydroxides consist of cationic metal hydroxide layers and exchangeable interlayer anions. This negatively charged structure permits biomolecules such as DNA to adsorb, and a so-called DNA-LDH hybrid is formed. The hydroxide layers can be removed in acidic media and the DNA will be released. CERATOFIX® (a registered trademark of SüdChemie AG NA that belongs to the family of LDHs, produced by Süd-Chemie AG). The chemical structure can be summarized as [Mg 2Al(OH)6](CO3)0.5. The binding capacity and kinetic characteristics of different types of CERATOFIX®NA for a model DNA was determined by flow cytometry. Results: The static binding capacities of the different LDHs were determined after 1- and 16-h incubation with DNA solution, showing different binding patterns between the LDH materials. The binding kinetics were revealed by flow cytometric measurements in short-term and long-term kinetic experiments, showing that the majority of DNA adsorbs within the first 60 s. Conclusions: DNA removal from cell culture supernatants is one of the major concerns in downstream processing. Due to the anion exchange capabilities of LDHs it seemed a very interesting approach to use these materials for binding of DNA for elimination purposes.
AB - Background: A new method was established to characterize the binding kinetics of DNA toward layered double hydroxides (LDHs). The setup consisted of a newly developed sampling tube that allows the injection of analyte during the flow cytometric measurement. Methods: Layered double hydroxides consist of cationic metal hydroxide layers and exchangeable interlayer anions. This negatively charged structure permits biomolecules such as DNA to adsorb, and a so-called DNA-LDH hybrid is formed. The hydroxide layers can be removed in acidic media and the DNA will be released. CERATOFIX® (a registered trademark of SüdChemie AG NA that belongs to the family of LDHs, produced by Süd-Chemie AG). The chemical structure can be summarized as [Mg 2Al(OH)6](CO3)0.5. The binding capacity and kinetic characteristics of different types of CERATOFIX®NA for a model DNA was determined by flow cytometry. Results: The static binding capacities of the different LDHs were determined after 1- and 16-h incubation with DNA solution, showing different binding patterns between the LDH materials. The binding kinetics were revealed by flow cytometric measurements in short-term and long-term kinetic experiments, showing that the majority of DNA adsorbs within the first 60 s. Conclusions: DNA removal from cell culture supernatants is one of the major concerns in downstream processing. Due to the anion exchange capabilities of LDHs it seemed a very interesting approach to use these materials for binding of DNA for elimination purposes.
KW - DNA removal
KW - Flow cytometry
KW - LDH
UR - http://www.scopus.com/inward/record.url?scp=8144230247&partnerID=8YFLogxK
U2 - 10.1002/cyto.a.20085
DO - 10.1002/cyto.a.20085
M3 - Article
C2 - 15478124
AN - SCOPUS:8144230247
VL - 62A
SP - 65
EP - 69
JO - Cytometry Part A
JF - Cytometry Part A
SN - 1552-4922
IS - 1
ER -