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

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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 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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Abbreviations

AEX:

Anion exchange

dNTP:

Deoxynucleotide

dsDNA:

Double-stranded DNA

HIC:

Hydrophobic interaction chromatography

IgG:

Immunoglobulin

MMC:

Multimodal chromatography

nt:

Nucleotide

pDNA:

Plasmid DNA

poly(dA):

Polydeoxyadenylate

poly(dC):

Polydeoxycytidylate

poly(dG):

Polydeoxyguanylate

poly(dT):

Polythymidylate

ssDNA:

Single-stranded DNA

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Acknowledgments

Tiago Matos acknowledges a fellowship (SFRH/BD/47934/2008) from FCT, Portuguese Foundation for Science and Technology. This work was supported by the Swedish Research Council (VR).

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Authors and Affiliations

  1. Department of Pure and Applied Biochemistry, Chemical Center, Lund University, P.O. Box 124, 22100, Lund, Sweden

    Tiago Matos, Elsayed T. Mohamed & Leif Bülow

  2. CICS-Health Sciences Research Centre, University of Beira Interior, 6201-001, Covilhã, Portugal

    Tiago Matos & João A. Queiroz

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  1. Tiago Matos
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  2. Elsayed T. Mohamed
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  3. João A. Queiroz
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Correspondence to Tiago Matos.

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Conflict of Interest

Tiago Matos was funded with fellowship (SFRH/BD/47934/2008) from FCT, Portuguese Foundation for Science and Technology, and declare no conflict of interest. This study was funded by Swedish Research Council (VR). All authors declare no conflict of interest of any type.

This article does not contain any studies with animals or with human participants performed by any of the authors.

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Matos, T., Mohamed, E.T., Queiroz, J.A. et al. Capto™ Resins for Chromatography of DNA: A Minor Difference in Ligand Composition Greatly Influences the Separation of Guanidyl-Containing Fragments. Chromatographia 79, 1277–1282 (2016). https://doi.org/10.1007/s10337-016-3148-3

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