Modified avidin and streptavidin molecules and use thereof

Details Modified avidin and streptavidin molecules and use thereof Modified avidin and streptavidin molecules and use thereof

C - Chemistry, Metallurgy

07

K

C07K 14/465 (2006.01) C07K 14/36 (2006.01) C12N 11/02 (2006.01) C12Q 1/68 (2006.01) G01N 33/53 (2006.01) G01N 33/531 (2006.01) G01N 33/543 (2006.01) G01N 33/566 (2006.01)

Patent

CA 2221647

Biotin-binding modified avidin-type molecules are provided in which the essential tyrosine residue in the bioting-binding site is modified in such a way that its pKa is decreased in comparison to the pKa of the unmodified tyrosine residue in the corresponding unmodified avidin-type molecule. The avidin-type molecules include: (i) native egg-white avidin; (ii) recombinant avidin; (iii) deglycosylated forms of avidin; (iv) bacterial streptavidin; (v) recombinant streptavidin; (vi) truncated streptavidin; and (vii) derivatives of (i)-(vi) which are modified at sites other than the essential tyrosine residue. The modification is achieved by substitution at one or both ortho positions to the hydroxy radical of the tyrosine residue by radicals such as nitro, halogen, azo and amino. The modified avidin-type molecules can be used in all applications of the avidin-biotin technology.

Molécules modifiées de type avidine liant la biotine, dans lesquelles le reste tyrosine essentiel dans le site de liaison de biotine est modifié de telle manière que son pKa est réduit par rapport à celui du reste tyrosine non modifié dans la molécule de type avidine non modifiée correspondante. Les molécules de type avidine comprennent (i) l'avidine native du blanc d'oeuf; (ii) l'avidine recombinante; (iii) les formes déglycosylées de l'avidine; (iv) la streptavidine bactérienne; (v) la streptavidine recombinante; (vi) la streptavidine tronquée; et (vii) des dérivés de (i) à (iv) modifiés au niveau de sites autres que le reste tyrosine essentiel. La modification s'effectue par substitution, au niveau de l'une et/ou l'autre de ses positions ortho, du radical hydroxyle du reste tyrosine par des radicaux tels que nitro, halogène, azo et amino. Ces molécules modifiées de type avidine sont utilisables dans toutes les applications mettant en oeuvre la technologie avidine-biotine.

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