Claims
- 1. A conjugate comprising a protease moiety conjugated to one or more polymers, wherein the protease moiety is a Bacillus sp. protease comprising one or more of the following substitutions: P6K, Y7K, S9K, A10K, Y11K, Q12E, Q12D, Q12K, R28K, Q33E, Q33D, D43K, Y44K, N45K, R51K, R62K, N65K, D68N, D68Q, G87K, I88K, D106N, D106Q, R121K, Q126D, Q126E, R169K, R19K, N209D, N209E, N209K, A211K, N216K, N217K, G218K, Y219K, S220K, Y221K, Q242D, Q242E, N245D, N245E, N246D, N246E, Q248D, Q248E, R250K, G262K, N266D and N266E.
- 2. The conjugate of claim 1, wherein the protease comprises one of the following substitutions: R28K+R62K, 28K+R169K, R62K+R169K, R28K+R69K+R169K.
- 3. The conjugate of claim 1, wherein the protease has an amino acid sequence of SEQ ID NO: 2.
- 4. The conjugate of claim 1, wherein the polymer(s) have a molecular weight from 1 to 60 kDa.
- 5. The conjugate of claim 1, wherein the polymer(s) are natural or synthetic homo- or heteropolymers.
- 6. The conjugate of claim 1, wherein the polymer(s) are selected from the group consisting of polyols, polyamines, polycarboxyl acids and polymers comprising a hydroxyl group and an amine group.
- 7. The conjugate of claim 1, wherein the polymer(s) are selected from the group consisting of polyalkylene oxides (PAO), PEG-glycidyl ethers (Epox-PEG), PEG-oxycarbonylimidazole (CDI-PEG), branched PEGs, poly-vinyl alcohols (PVA), poly-carboxylates, polyvinylpyrolidones, poly-D,L-amino acids, polyethylene-co-maleic acid anhydride, polystyrene-co-malic acid anhydrides, dextrans, heparins, homologous albumins, celluloses, hydrolysates of chitosan, starches, glycogen, agaroses and derivatives thereof, guar gum, pullulan, inulin, xanthan gum, carrageenin, pectin, alginic acid hydrolysates and bio-polymers.
- 8. The conjugate of claim 1, wherein the polymer(s) are polyalkylene glycols (PAG) or methoxypolyethylene glycols (mPEG).
- 9. The conjugate of claim 1, wherein the polymer(s) are selected from the group consisting of polyethylene glycols (PEG), polypropylene glycols and carboxymethyl-dextrans.
- 10. The conjugate of claim 1, wherein the polymer(s) are selected from the group consisting of methylcellulose, carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose carboxyethylcellulose and hydroxypropylcellulose.
- 11. The conjugate of claim 1, wherein the polymer(s) are hydroxyethyl-starches or hydroxypropyl-starches.
- 12. The conjugate of claim 1, wherein the polymer(s) are methoxypolyethylene glycols (mPEG).
- 13. A conjugate comprising a peroxidase moiety conjugated to one or more polymers, wherein the peroxidase moiety is an Arthromyces ramosus peroxidase comprising one or more of the following substitutions: Q34D, Q34E, Q70D, Q70E, N128D, N128E, N303D and N303E.
- 14. The conjugate of claim 13, wherein the peroxidase has an amino acid sequence of SEQ ID NO: 4.
- 15. The conjugate of claim 13, wherein the polymer(s) have a molecular weight from 1 to 60 kDa.
- 16. The conjugate of claim 13, wherein the polymer(s) are natural or synthetic homo- or heteropolymers.
- 17. The conjugate of claim 13, wherein the polymer(s) are selected from the group consisting of polyols, polyamines, polycarboxyl acids and polymers comprising a hydroxyl group and an amine group.
- 18. The conjugate of claim 13, wherein the polymer(s) are selected from the group consisting of polyalkylene oxides (PAO), PEG-glycidyl ethers (Epox-PEG), PEG-oxycarbonylimidazole (CDI-PEG), branched PEGs, poly-vinyl alcohols (PVA), poly-carboxylates, polyvinylpyrolidones, poly-D,L-amino acids, polyethylene-co-maleic acid anhydride, polystyrene-co-malic acid anhydrides, dextrans, heparins, homologous albumins, celluloses, hydrolysates of chitosan, starches, glycogen, agaroses and derivatives thereof, guar gum, pullulan, inulin, xanthan gum, carrageenin, pectin, alginic acid hydrolysates and bio-polymers.
- 19. The conjugate of claim 13, wherein the polymer(s) are polyalkylene glycols (PAG) or methoxypolyethylene glycols (mPEG).
- 20. The conjugate of claim 13, wherein the polymer(s) are selected from the group consisting of polyethylene glycols (PEG), polypropylene glycols and carboxymethyl-dextrans.
- 21. The conjugate of claim 13, wherein the polymer(s) are selected from the group consisting of methylcellulose, carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose carboxyethylcellulose and hydroxypropylcellulose.
- 22. The conjugate of claim 13, wherein the polymer(s) are hydroxyethyl-starches or hydroxypropyl-starches.
- 23. The conjugate of claim 13, wherein the polymer(s) are methoxypolyethylene glycols (mPEG).
Priority Claims (1)
Number |
Date |
Country |
Kind |
0135/97 |
Feb 1997 |
DK |
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CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of application serial No. PCT/DK98/00046 filed on Feb. 6, 1998 and claims priority under 35 U.S.C. 119 of Danish application serial No. 0135/97 filed Feb. 6, 1997, the contents of which are fully incorporated herein by reference.
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Continuations (1)
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Number |
Date |
Country |
Parent |
PCT/DK98/00046 |
Feb 1998 |
US |
Child |
09/024532 |
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US |