DOI: 10.1111/j.1432-1033.1981.tb05060.x. Receptor‐Specific Large‐Scale Purification of Cholera Toxin on Silica Beads Derivatized with LysoG<sub>M1</sub> Ganglioside

Receptor‐Specific Large‐Scale Purification of Cholera Toxin on Silica Beads Derivatized with LysoGM1 Ganglioside

10.1111/j.1432-1033.1981.tb05060.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

A receptor‐specific affinity chromatographic method for large‐scale purification of cholera toxin is described. The receptor ganglioside for cholera toxin, GM1, is hydrolysed to lysoGM1 which is then covalently coupled, via stabilized Schiff's bases, to porous silica beads (Spherosil) onto which a layer of DEAE‐dextran has been adsorbed and cross‐linked before coupling. Columns of these Spherosil‐DEAE‐dextran‐lysoGM1 beads, in contrast to particles derivatized with lysoGA1, bound the cholera toxin of Vibrio cholerae culture filtrates, after which the toxin could be eluted with the aid of an acid citrate buffer (pH 2.8). The toxin‐binding capacity was directly proportional to the amount of lysoGM1 in the column: 2.3 mg/μmol lysoGM1. The yield of purified toxin after acid elution and pH neutralization was essentially quantitative (83–107%). The affinity‐purified toxin contained less than 5% impurities, but consisted of a mixture of predominantly intact holotoxin and B subunit protomer which could readily be separated by gel filtration on Sephadex G‐100. Scaling up of the technique was possible: a 1 kg column enabled us to treat 1000‐l cultures of V. cholerae and thus to isolate 20 g of cholera toxin per cycle.

Bibliography

TAYOT, J., HOLMGREN, J., SVENNERHOLM, L., LINDBLAD, M., & TARDY, M. (1981). ReceptorâSpecific LargeâScale Purification of Cholera Toxin on Silica Beads Derivatized with LysoGM1 Ganglioside. European Journal of Biochemistry, 113(2), 249â258. Portico.

Authors 5

Jean‐Louis TAYOT (first)
Jan HOLMGREN (additional)
Lars SVENNERHOLM (additional)
Marianne LINDBLAD (additional)
Michel TARDY (additional)

References 25 Referenced 68

{'key': 'e_1_2_2_2_2', 'first-page': '85', 'volume': '8', 'author': 'Gill D. M.', 'year': '1977', 'journal-title': 'Adv. Cyclic Nucleotide Res.'} / Adv. Cyclic Nucleotide Res. by Gill D. M. (1977)
10.1111/j.1469-185X.1977.tb00858.x
{'key': 'e_1_2_2_4_2', 'first-page': '269', 'volume-title': 'The Molecular Basis of Microbiol Pathogenicity', 'author': 'Holmgren J.', 'year': '1980'} / The Molecular Basis of Microbiol Pathogenicity by Holmgren J. (1980)
{'key': 'e_1_2_2_5_2', 'first-page': '88', 'volume-title': 'Cholera and Related Diarrheas', 'author': 'Holmgren J.', 'year': '1980'} / Cholera and Related Diarrheas by Holmgren J. (1980)
10.1146/annurev.bi.48.070179.003053
10.1093/infdis/136.Supplement.S105
{'key': 'e_1_2_2_8_2', 'first-page': '171', 'volume-title': 'Cholera and Related Diarrheas', 'author': 'Svennerholm A. M.', 'year': '1980'} / Cholera and Related Diarrheas by Svennerholm A. M. (1980)
10.1038/269602a0
{'key': 'e_1_2_2_10_2', 'first-page': '95', 'volume-title': 'Chromatography of Synthetic and Biological Polymers', 'author': 'Tavot J. ‐L.', 'year': '1978'} / Chromatography of Synthetic and Biological Polymers by Tavot J. ‐L. (1978)
10.1016/0005-2760(80)90052-1
10.1016/0009-8981(79)90396-6
10.1016/0006-3002(57)90254-8
10.1007/978-1-4684-7844-0_35
10.1016/0019-2791(65)90004-2
10.1007/BF02601736
10.1128/iai.7.5.759-763.1973
10.1128/iai.8.6.851-859.1973
10.1007/BF02601701
10.1099/00221287-76-2-417
10.1038/207614a0
10.1128/IAI.8.2.208-214.1973
{'key': 'e_1_2_2_23_2', 'first-page': '229', 'volume': '52', 'author': 'Holmgren J.', 'year': '1974', 'journal-title': 'Med. Biol. (Helsinki)'} / Med. Biol. (Helsinki) by Holmgren J. (1974)
10.1016/S0076-6879(74)34009-8
10.1016/0022-1759(76)90107-1
10.1021/bi00745a033

Dates

Type	When
Created	20 years, 6 months ago (March 3, 2005, 3:57 p.m.)
Deposited	1 year, 9 months ago (Nov. 22, 2023, 7:34 p.m.)
Indexed	1 day, 6 hours ago (Sept. 4, 2025, 9:46 a.m.)
Issued	44 years, 8 months ago (Jan. 1, 1981)
Published	44 years, 8 months ago (Jan. 1, 1981)
Published Online	20 years, 6 months ago (March 3, 2005)
Published Print	44 years, 8 months ago (Jan. 1, 1981)

Funders 0

None

BibTeX

@article{TAYOT_1981, title={Receptor‐Specific Large‐Scale Purification of Cholera Toxin on Silica Beads Derivatized with LysoGM1 Ganglioside}, volume={113}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1981.tb05060.x}, DOI={10.1111/j.1432-1033.1981.tb05060.x}, number={2}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={TAYOT, Jean‐Louis and HOLMGREN, Jan and SVENNERHOLM, Lars and LINDBLAD, Marianne and TARDY, Michel}, year={1981}, month=jan, pages={249–258} }

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