Claims
- 1. An apparatus for conducting a plurality of microscale reactions, comprising
a. a solid support comprising a first and a second reaction site, wherein each site comprises a distinct immobilized biocatalyst; b. a micro-directing element to direct an intermediate composition from the first site to the second site.
- 2. The apparatus of claim 1, further comprising a micro-directing element to direct a composition from a source to at least one said reaction site.
- 3. The apparatus of claim 2, wherein each said biocatalyst is a catalytic antibody, an enzyme, an enzyme domain, a multi-enzyme complex, a catalytic peptide or a catalytic RNA molecule.
- 4. The apparatus of claim 2, wherein
a. the apparatus comprises n additional reaction sites; b. n is an integer from 1 to 50; c. each reaction site comprises a distinct immobilized biocatalyst; and d. the apparatus additionally comprises one or more micro-directing elements to sequentially direct a composition from the second reaction site to each of the n additional reaction sites.
- 5. The apparatus of claim 3, wherein the apparatus additionally comprises at least one reaction site which comprises a catalyst of non-biological origin.
- 6. The apparatus of claim 3, wherein the biocatalyst at the first reaction and the biocatalyst at the second reaction site are each a functional catalytic unit from sequential steps in a natural biosynthetic pathway.
- 7. The apparatus of claim 4, wherein:
a. the biocatalyst at the first reaction site, the second reaction site and each of the n additional reaction sites are a functional catalytic unit from the steps of a natural biosynthetic pathway; b. the apparatus additionally comprises one or more micro-directing elements to direct a composition sequentially to each of the reaction sites in the order of the sequence of the catalytic units in the biosynthetic pathway.
- 8. The apparatus of claim 7, wherein said biosynthetic pathway is the picromycin polyketide synthase pathway, the erythromycin polyketide synthase pathway, the nonribosomal peptide synthetase pathway, or the β-lactam nonribosomal peptide synthetase pathway.
- 9. The apparatus of claim 4, wherein each biocatalyst is immobilized by a material comprising, a polyacrylamide, a polyacrylate, a sugar-substituted polyacrylate, a polyvinyl alcohol, a polyvinylene, or a polyvinyl silicate.
- 10. The apparatus of claim 9, wherein each said material comprises a polyacrylate substituted with sucrose, lactose, mannose, an alkylmannoside, glucose, an alkylglucoside, galactose, an alkylgalactoside, fructose, an alkylfructoside, or trehalose.
- 11. The apparatus of claim 10, wherein said micro-directing elements comprise a channel or a conduit between each reaction site in the sequence.
- 12. The apparatus of claim 11, wherein said apparatus additionally comprising a device to propel a fluid between the reaction sites using electroosmotic force.
- 13. The apparatus of claim 12, further comprising a device to control the temperature of at least a portion of the apparatus.
- 14. The apparatus of claim 13, further comprising a micro-directing element to direct at least one said intermediate composition from one said reaction site to a detector.
- 15. The apparatus of claim 14, further comprising a detector, wherein said detector comprises an aspiration probe, a laser desorption probe, an ion beam desorption probe, a gas desorption probe, a liquid desorption probe, a contact probe, an optical spectrometer, a microscope, an imager, a mass spectrometer, a chromatography apparatus, an electrochemical detector, a particle detector, a radiation detector or a magnetic resonance spectrometer.
- 16. A microfluidics chip, comprising
a. a solid support comprising a first and second reaction site, wherein
i. the first and second reaction sites each comprise a distinct biocatalyst that is a functional catalytic unit of a natural biosynthetic pathway; ii. each said biocatalyst is immobilized by a polyacrylate substituted with sucrose, lactose, mannose, an alkylmannoside, glucose, an alkylglucoside, galactose, an alkylgalactoside, fructose, an alkylfructoside, or trehalose; b. directing a substrate composition by microfluidics from a source to the first reactive composition, and directing an intermediate composition by microfluidics from the first reaction site to the second reaction site, wherein the biocatalysts at the first and second sites are from sequential steps in the natural biosynthetic pathway
- 17. A method for conducting reactions at a plurality of reaction sites, comprising the steps of:
a. providing an apparatus, wherein
i. said apparatus is a solid support comprising a first reaction site and a second reaction site; ii. each reaction site comprises a distinct immobilized biocatalyst; and b. directing a substrate composition to said first reaction site and exposing the first reaction site to conditions suitable for reacting said substrate with said immobilized catalyst, thereby producing a first intermediate composition; and c. directing said first intermediate composition to said second reaction site and exposing the second reaction site to conditions suitable for reacting said first intermediate with said immobilized catalyst, thereby producing a second intermediate composition.
- 18. The method of claim 17, further comprising the step of directing a composition from a source to at least one said reaction site.
- 19. The method of claim 17, wherein
a. the apparatus additionally comprises n additional reaction sites; b. n is an integer from 1 to 50; c. each reaction site comprises a distinct immobilized biocatalyst; and d. the apparatus additionally comprises one or more micro-directing elements to sequentially direct a composition from the second reaction site to each of the n additional reaction sites, wherein the method further comprises the steps of:
i. directing the second intermediate to the next reaction site in the sequence and exposing the next reaction site to conditions suitable for reacting the second intermediate with the biocatalyst at the next site, thereby producing a third intermediate; ii. repeating step (i) with the third intermediate and each subsequent intermediate.
- 20. The method of claim 19, wherein each said biocatalyst is a catalytic antibody, an enzyme, an enzyme domain, a multi-enzyme complex, a catalytic peptide or a catalytic RNA molecule.
- 21. The method of claim 17, wherein the substrate is an unnatural substrate for the biocatalyst at the first reaction site, or one of the intermediates is an unnatural substrate for the biocatalyst at the next reaction site in the sequence.
- 22. The method of claim 17, wherein the apparatus additionally comprises a catalyst of non-biological origin.
- 23. The method of claim 17, wherein the biocatalyst at the first reaction site and at the second reaction site are each a distinct functional catalytic unit from sequential steps in a natural biosynthetic pathway.
- 24. The method of claim 23, wherein said biosynthetic pathway is the picromycin polyketide synthase pathway, the erythromycin polyketide synthase pathway, the nonribosomal peptide synthetase pathway, or the β-lactam nonribosomal peptide synthetase pathway.
- 25. The method of claim 19, wherein:
a. the biocatalyst at the first reaction site, the second reaction site and each of the n additional reaction sites are a functional catalytic unit from the steps of a natural biosynthetic pathway; b. wherein the reaction sequence of the apparatus and the reaction sequence of the biosynthetic pathway are the same.
- 26. The method of claim 17, wherein each said immobilized catalyst is immobilized by a material comprising a polyacrylamide, a polyacrylate, a sugar-substituted polyacrylate, a polyvinyl alcohol, a polyvinylene, or a polyvinyl silicate.
- 27. The method of claim 26, wherein each said material comprises a polyacrylate substituted with a sugar comprising sucrose, lactose, mannose, an alkylmannoside, glucose, an alkylglucoside, galactose, an alkylgalactoside, fructose, an alkylfructoside, or trehalose.
- 28. The method of claim 27, wherein the micro-directing element is a channel or a conduit between each reaction site in the sequence; and the compositions are directed by pressure, electroosmotic force, piezoelectric force, electrostatic force, centrifugal force, or a force generated by centripetal acceleration.
- 29. The method of claim 28, further comprising the step of directing said compositions by electroosmotic force.
- 30. The method of claim 29, further comprising the step of detecting a constituent of a composition.
- 31. The method of claim 30, wherein the step of detecting employs an aspiration probe, a laser desorption probe, an ion beam desorption probe, a gas desorption probe, a liquid desorption probe, a contact probe, an optical spectrometer, a microscope, an imager, a mass spectrometer, a chromatography apparatus, an electrochemical detector, a particle detector, a radiation detector or a magnetic resonance spectrometer.
- 32. The method of claim 31, further comprising the step of controlling the temperature.
- 33. A method for conducting sequential reactions, comprising the steps of:
a. providing a microfluidics chip, wherein:
i. said chip comprises a first reaction site and a second reaction site; and ii. each said reaction site comprises an immobilized biocatalyst, wherein each said catalyst is a catalytic antibody, an enzyme, an enzyme domain, a multi-enzyme complex, a catalytic peptide or a catalytic RNA molecule; and b. directing a substrate composition to the first reaction site under conditions suitable for reacting said substrate with said biocatalyst, thereby producing a first intermediate composition; and c. directing said first intermediate to the second reaction site under conditions suitable for reacting said intermediate with said biocatalyst, thereby producing a second intermediate composition.
- 34. The method of claim 33 wherein the biocatalysts in step (ii) combined with step (b) and step (c) collectively constitute the sequence of a portion of a natural biosynthetic pathway.
- 35. The method of claim 33 wherein the biocatalysts in step (ii) combined with step (b) and step (c) collectively constitute an unnatural sequence.
- 36. The apparatus of claim 1, wherein each biocatalyst is immobilized by a material, wherein said material comprises a solgel, a collagen gel, a hydrogel, or a polysaccharide gel
- 37. The apparatus of claim 36, wherein a solgel is substituted or unsubstituted and is a tetramethoxyorthosilicate, a methyl-trimethoxyorthosilicate, a tetraalkoxyorthosilicate, or a trialkoxyorthosilicate.
- 38. The apparatus of claim 37, wherein a hydrogel is substituted or unsubstituted and is a polyacrylamide, a polyacrylate, a sugar-substituted polyacrylate, or a polyvinyl alcohol.
- 39. The apparatus of claim 38, wherein a polysaccharide gel is substituted or unsubstituted and is an alginate, a dextran, a starch, a cellulose, a carrageenan, a poly(hyaluronic acid), a heparin, a guar, or an inulin.
RELATED APPLICATION
[0001] This application claims the benefit of U.S. Provisional Application No. 60/336,045, filed on Nov. 1, 2001, the entire teachings of which are incorporated herein by reference.
GOVERNMENT SUPPORT
[0002] The invention was supported, in whole or in part, by a grant from the Office of Naval Research and the National Institutes of Health. The Government has certain rights in the invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60336045 |
Nov 2001 |
US |