Claims
- 1. A composition comprising an isolated nucleic acid having the sequence of SEQ ID NO: 1.
- 2. A vector comprising the nucleic acid of claim 1.
- 3. The vector of claim 2, wherein said vector is an expression vector.
- 4. The expression vector of claim 3, wherein said nucleic acid sequence is operably linked to an inducible promoter.
- 5. A cell comprising the expression vector of claim 3.
- 6. The cell of claim 5, wherein said cell is a bacterial cell.
- 7. The cell of claim 6, wherein said cell further comprises the protein encoded by said nucleic acid sequence.
- 8. A method, comprising:
a) providing:
(i) M. oreades mushrooms, and (ii) an extraction buffer comprising a protease inhibitor cocktail and ethylenediaminetetraacetic acid; b) preparing an extract of said M. oreades mushrooms in said extraction buffer; and c) treating said extract under conditions such that a lectin preparation is produced.
- 9. The method of claim 8, wherein said preparing step b) is carried out under an atmosphere of argon.
- 10. The method of claim 8, wherein said treating step c) comprises column chromatography.
- 11. The method of claim 10, wherein said column chromatography comprises a melibiose-Sepharose affinity column.
- 12. The method of claim 8, comprising an additional purification step d) passing said lectin preparation through an affinity column to produce a purified lectin.
- 13. The method of claim 12, wherein said affinity column of step d) comprises Galα1,3 [L-Fucα1,2]GalβO(CH2)8CONH.
- 14. The method of claim 12, wherein said lectin has high binding affinity for Galα1,3Gal.
- 15. The method of claim 8, wherein said lectin preparation is substantially free of proteolytic fragments of said lectin.
- 16. The method of claim 8, wherein said binding affinity is determined by a method selected from the group consisting of isothermal titration calorimetry, blood cell agglutination and carbohydrate precipitation.
- 17. A composition comprising the purified lectin of claim 12.
- 18. A method, comprising:
a) providing:
(i) a composition comprising labeled M. oreades agglutinin, and (ii) a sample obtained from a subject; and b) contacting said sample with said labeled M. oreades agglutinin under conditions such that a glycoconjugate in said sample is detected.
- 19. The method of claim 18, wherein said subject has one or more symptoms of malaria.
- 20. The method of claim 18, wherein said sample is a human blood sample.
- 21. The method of claim 20, wherein said human blood sample comprises Plasmodium falciparum merozoites.
- 22. The method of claim 18, wherein said labeled M. oreades agglutinin comprises a fluorescent label.
- 23. The method of claim 22, wherein said fluorescent label is fluorescein.
- 24. A composition comprising immobilized M. oreades agglutinin.
- 25. The composition of claim 24, wherein said immobilized M. oreades agglutinin is immobilized on Sepharose 4B.
- 26. The composition of claim 24, wherein said Sepharose 4B-immobilized M. oreades agglutinin is contained in a column.
- 27. A method, comprising:
a) providing:
i) an immobilized-M. oreades agglutinin affinity column, ii) a sample suspected of comprising a saccharide selected from the group consisting of Galα1,3Gal, Galα1,3Galβ1,4GlcNAc and Galα1,3 [L-Fucα1,2]Gal; b) contacting said sample with said affinity column under conditions such that said saccharide is bound in a complex; and c) treating said complex under conditions such that said saccharide is released from said complex.
- 28. The method of claim 27, wherein said sample is a serum sample.
PRIORITY
[0001] This application claims the benefit of U.S. Provisional Application Serial No. 60/288,596, filed on May 3, 2001 and U.S. Provisional Application Serial No. 60/354,322, filed on Feb. 4, 2002.
STATEMENT OF GOVERNMENTAL INTEREST
[0002] This invention was made with government support awarded by the National Institutes of Health under grant number GM 29470-35. The Government has certain rights in the invention.
Provisional Applications (2)
|
Number |
Date |
Country |
|
60288596 |
May 2001 |
US |
|
60354322 |
Feb 2002 |
US |