Muteins of bilin-binding protein

Details Muteins of bilin-binding protein Muteins of bilin-binding protein

2006-02-21

2006-02-21

Carlson, Karen Cochrane (Department: 1653)

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Peptide containing doai

C514S002600, C530S350000, C530S395000, C530S378000, C435S069700, C435S440000

Reexamination Certificate

active

07001882

ABSTRACT:
The present invention relates to muteins of the bilin-binding protein with binding activity to digoxigenin and to fusion proteins of such muteins, a method for preparing said muteins and fusion proteins thereof and their utilization for detecting or binding digoxigenin-labeled biomolecules. The invention especially relates to a polypeptide selected from muteins of the bilin-binding protein, characterized in that (a) it can bind digoxigenin or digoxigenin conjugates, (b) it does not bind ouabain, testosterone, and 4-aminofluorescein and (c) at least one of the sequence positions 28, 31, 34, 35, 36, 37, 58, 60, 69, 88, 90, 95, 97, 114, 116, 125, and 127 of the bilin-binding protein has an amino acid substitution. Due to their simple molecular structure, the inventive muteins provide advantages for production and utilization in comparison with antibodies against the digoxigenin group.

REFERENCES:
patent: 0 835 934 (1998-04-01), None
patent: 89/06698 (1989-07-01), None
patent: 99/16873 (1999-04-01), None
Beste et al., “Small Antibody-Like Proteins with Prescribed Ligand Specificities Derived From the Lipocalin Fold,” Biochemistry, Proceeding of the National Academy of Science of the United States, vol. 96, No. 5, pp. 1898-1903, Mar. 2, 1999.
Flower, “The Lipocalin Protein Family: Structure and Function,” Biochemical Journal, vol. 318, pp. 1-14, 1996.
Schmidt et al, “The Bilin-Binding Protein ofPieris brassicaecDNA Sequence and Regulation of Expression Reveal Distinct Features of this Insect Pigment Protein,” European Journal of Biochemistry, vol. 219, No. 3, FEBS, pp. 855-863, 1994.
Schlehuber et al., “A Novel Type of Receptor Protein, Based on the Lipocalin Scaffold, with Specificity for Digoxigenin,” Journal of Molecular Biology, vol. 297, No. 5, Academic Press, pp. 1105-1120, Apr. 14, 2000.

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