High-performance liquid affinity chromatography on silica-bound alcohol dehydrogenase
(1983) In Analytical Biochemistry 134(1). p.60-72- Abstract
- Horse liver alcohol dehydrogenase was immobilized on glycerylpropyl-silica (10 μm, 1000-Å pores) activated with 2,2,2-trifluoroethanesulfonyl chloride (tresyl chloride). The coupling and activity yield was almost 100%. The coenzyme-binding sites were equivalent and virtually unaffected by the immobilization process, as judged from Scatchard plots and active-site titrations. The silica-bound enzyme, packed in steel columns, was integrated with HPLC equipment and then successfully used for chromatography of adenine nucleosides, adenine nucleotides, and triazine dyes. Dissociation constants were calculated from chromatographic data and found to correspond well with literature values. The dissociation constants for a number of nucleotide... (More)
- Horse liver alcohol dehydrogenase was immobilized on glycerylpropyl-silica (10 μm, 1000-Å pores) activated with 2,2,2-trifluoroethanesulfonyl chloride (tresyl chloride). The coupling and activity yield was almost 100%. The coenzyme-binding sites were equivalent and virtually unaffected by the immobilization process, as judged from Scatchard plots and active-site titrations. The silica-bound enzyme, packed in steel columns, was integrated with HPLC equipment and then successfully used for chromatography of adenine nucleosides, adenine nucleotides, and triazine dyes. Dissociation constants were calculated from chromatographic data and found to correspond well with literature values. The dissociation constants for a number of nucleotide derivatives with potential application in affinity chromatography were also determined. The spacers were found to affect the binding strength of the nucleotides in a qualitatively predictable way. Theoretical plate heights were calculated and found to be in the range 0.01 to 0.1 cm. Attempts to correlate peak widths with the rate constants for the binary complexes involved were only partially successful. (Less)
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https://lup.lub.lu.se/record/35351063-fa6b-4cb3-a6cb-d28ed223a523
- author
- Nilsson, Kurt and Larsson, Per-Olof LU
- organization
- publishing date
- 1983
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Analytical Biochemistry
- volume
- 134
- issue
- 1
- pages
- 13 pages
- publisher
- Elsevier
- external identifiers
-
- scopus:0020838007
- ISSN
- 0003-2697
- DOI
- 10.1016/0003-2697(83)90264-6
- language
- English
- LU publication?
- yes
- id
- 35351063-fa6b-4cb3-a6cb-d28ed223a523
- date added to LUP
- 2024-06-18 18:38:44
- date last changed
- 2024-09-06 14:50:04
@article{35351063-fa6b-4cb3-a6cb-d28ed223a523,
abstract = {{Horse liver alcohol dehydrogenase was immobilized on glycerylpropyl-silica (10 μm, 1000-Å pores) activated with 2,2,2-trifluoroethanesulfonyl chloride (tresyl chloride). The coupling and activity yield was almost 100%. The coenzyme-binding sites were equivalent and virtually unaffected by the immobilization process, as judged from Scatchard plots and active-site titrations. The silica-bound enzyme, packed in steel columns, was integrated with HPLC equipment and then successfully used for chromatography of adenine nucleosides, adenine nucleotides, and triazine dyes. Dissociation constants were calculated from chromatographic data and found to correspond well with literature values. The dissociation constants for a number of nucleotide derivatives with potential application in affinity chromatography were also determined. The spacers were found to affect the binding strength of the nucleotides in a qualitatively predictable way. Theoretical plate heights were calculated and found to be in the range 0.01 to 0.1 cm. Attempts to correlate peak widths with the rate constants for the binary complexes involved were only partially successful.}},
author = {{Nilsson, Kurt and Larsson, Per-Olof}},
issn = {{0003-2697}},
language = {{eng}},
number = {{1}},
pages = {{60--72}},
publisher = {{Elsevier}},
series = {{Analytical Biochemistry}},
title = {{High-performance liquid affinity chromatography on silica-bound alcohol dehydrogenase}},
url = {{http://dx.doi.org/10.1016/0003-2697(83)90264-6}},
doi = {{10.1016/0003-2697(83)90264-6}},
volume = {{134}},
year = {{1983}},
}