The Purification of Biologically Active Compounds by Affinity Chromatography

Wilchek, Meir; Hexter, Charles S.

doi:10.1002/9780470110430.ch5

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B Biotin Affinity Columns1

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1979

1992

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Cited by 17 publications

(1 citation statement)

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“…The synthesis of poly-L-lysine or polyacrylic hydrazide and its subsequent coupling to Sepharose was THE USE OF THE AVIDIN-BIOTIN COMPLEX 15 described earlier in this series (Wilchek and Hexter, 1976). Poly-L-lysylSepharose (3 g, containing about 60 pmole of lysine) was suspended in absolute dioxane (3 ml).…”

Section: B Biotin Affinity Columnsmentioning

confidence: 99%

The Use of the Avidin‐Biotin Complex as a Tool in Molecular Biology

Bayer

Wilchek

1980

Methods of Biochemical Analysis

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551

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Section: B Biotin Affinity Columnsmentioning

confidence: 99%

The Use of the Avidin‐Biotin Complex as a Tool in Molecular Biology

Bayer

Wilchek

1980

Methods of Biochemical Analysis

Self Cite

551

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Glycosidases—Properties and Application to the Study of Complex Carbohydrates and Cell Surfaces

Flowers¹,

Sharon²

1979

Advances in Enzymology - And Related Areas of Molecular Biology

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Mechanism of reaction of cyanogen bromide‐activated agarose with amines and the solvolysis of amine ligands

Kennedy¹,

Barnes²,

Matthews³

1983

Brit. Poly. J.

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Cyanogen bromide activated agarose is widely employed as the water‐insoluble matrix in immunoadsorption and affinity chromatography for the immobilisation of proteins. A reaction mechanism is presented for the cyanogen bromide activation of the galactan, agarose, the coupling of model amines to the activated carbohydrate matrix and the detachment of ligands. Dansyl (Dns) derivatisation of the acid hydrolysed and unhydrolysed amine derivatives resulting from the reaction of cyanogen bromide‐activated Sepharose® and N‐α‐acetyl‐L‐ornithine reveal the production of arginine, indicating that the amine‐Sepharose conjugate is an O‐substituted isourea. Thin layer chromatography of the Dns‐derivatised unhydrolysed filtrate from the reaction indicated the formation of several derivatives which suggest that the isourea bonds can be cleaved by the solvolytic attack of nucleophiles in the pH range 8–9.5. Nucleophilic attack by amino groups on the amine‐Sepharose conjugates releases N‐monosubstituted guanidine, N1, N2 disubstituted ornithine and substituted ureas.

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The Purification of Biologically Active Compounds by Affinity Chromatography

Cited by 17 publications

References 253 publications

The Use of the Avidin‐Biotin Complex as a Tool in Molecular Biology

The Use of the Avidin‐Biotin Complex as a Tool in Molecular Biology

Glycosidases—Properties and Application to the Study of Complex Carbohydrates and Cell Surfaces

Mechanism of reaction of cyanogen bromide‐activated agarose with amines and the solvolysis of amine ligands

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