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Capto™ Resins for Chromatography of DNA : A Minor Difference in Ligand Composition Greatly Influences the Separation of Guanidyl-Containing Fragments

Matos, Tiago LU ; Mohamed, Elsayed T.; Queiroz, João A. and Bülow, Leif LU (2016) In Chromatographia 79(19-20). p.1277-1282
Abstract

The differences in chromatographic behaviour of individual deoxynucleotides as well as small single-stranded and double-stranded DNA molecules have been examined for two resins from the Capto family: Capto Adhere and Capto Q ImpRes. Capto Adhere carries a multimodal ligand which combines strong anion with aromatic recognition, while Capto Q ImpRes is a strong anion exchanger with a chemically similar ligand, but without a phenyl group. The intrinsic differences between single- and double-stranded DNAs are related to charge, hydrophobicity, size and three-dimensional structure. These variations in biophysical properties have been utilized for comparative separations on these two resins. All deoxynucleotides and DNAs tested bound strongly... (More)

The differences in chromatographic behaviour of individual deoxynucleotides as well as small single-stranded and double-stranded DNA molecules have been examined for two resins from the Capto family: Capto Adhere and Capto Q ImpRes. Capto Adhere carries a multimodal ligand which combines strong anion with aromatic recognition, while Capto Q ImpRes is a strong anion exchanger with a chemically similar ligand, but without a phenyl group. The intrinsic differences between single- and double-stranded DNAs are related to charge, hydrophobicity, size and three-dimensional structure. These variations in biophysical properties have been utilized for comparative separations on these two resins. All deoxynucleotides and DNAs tested bound strongly to the chromatographic materials and could be eluted by a linear gradient of increasing NaCl concentration. Capto Q ImpRes provided a recognition for guanylate bases when samples of deoxynucleotides or poly(dG) were examined. This recognition was not observed for Capto Adhere. Another pronounced difference between the resins was observed in the inverted elution of ss- and dsDNA, where ssDNA eluted at 2.88 M NaCl on Capto Adhere, while on Capto Q ImpRes ssDNA eluted already at 1.47 M NaCl. This behaviour can be linked to the presence of the more hydrophobic phenyl group in Capto Adhere, leading to stronger retention of ssDNA molecules, which have a more hydrophobic character due to a higher degree of base exposure.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Double-stranded DNA, Ion-exchange chromatography, Nucleotide, Oligodeoxynucleotide, Plasmid, Single-stranded DNA
in
Chromatographia
volume
79
issue
19-20
pages
6 pages
publisher
Vieweg Verlag
external identifiers
  • Scopus:84980007094
  • WOS:000387326400006
ISSN
0009-5893
DOI
10.1007/s10337-016-3148-3
language
English
LU publication?
yes
id
9a20ff31-e682-4655-8710-3532a455686a
date added to LUP
2016-10-17 10:33:32
date last changed
2017-01-01 08:37:00
@article{9a20ff31-e682-4655-8710-3532a455686a,
  abstract     = {<p>The differences in chromatographic behaviour of individual deoxynucleotides as well as small single-stranded and double-stranded DNA molecules have been examined for two resins from the Capto family: Capto Adhere and Capto Q ImpRes. Capto Adhere carries a multimodal ligand which combines strong anion with aromatic recognition, while Capto Q ImpRes is a strong anion exchanger with a chemically similar ligand, but without a phenyl group. The intrinsic differences between single- and double-stranded DNAs are related to charge, hydrophobicity, size and three-dimensional structure. These variations in biophysical properties have been utilized for comparative separations on these two resins. All deoxynucleotides and DNAs tested bound strongly to the chromatographic materials and could be eluted by a linear gradient of increasing NaCl concentration. Capto Q ImpRes provided a recognition for guanylate bases when samples of deoxynucleotides or poly(dG) were examined. This recognition was not observed for Capto Adhere. Another pronounced difference between the resins was observed in the inverted elution of ss- and dsDNA, where ssDNA eluted at 2.88 M NaCl on Capto Adhere, while on Capto Q ImpRes ssDNA eluted already at 1.47 M NaCl. This behaviour can be linked to the presence of the more hydrophobic phenyl group in Capto Adhere, leading to stronger retention of ssDNA molecules, which have a more hydrophobic character due to a higher degree of base exposure.</p>},
  author       = {Matos, Tiago and Mohamed, Elsayed T. and Queiroz, João A. and Bülow, Leif},
  issn         = {0009-5893},
  keyword      = {Double-stranded DNA,Ion-exchange chromatography,Nucleotide,Oligodeoxynucleotide,Plasmid,Single-stranded DNA},
  language     = {eng},
  month        = {10},
  number       = {19-20},
  pages        = {1277--1282},
  publisher    = {Vieweg Verlag},
  series       = {Chromatographia},
  title        = {Capto™ Resins for Chromatography of DNA : A Minor Difference in Ligand Composition Greatly Influences the Separation of Guanidyl-Containing Fragments},
  url          = {http://dx.doi.org/10.1007/s10337-016-3148-3},
  volume       = {79},
  year         = {2016},
}