Methods and compositions for the detection and/or quantification of gram positive bacterial contaminants
First Claim
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1. A method of producing a preparation for detecting the presence and/or amount of lipoteichoic acid in a sample, the method comprising the steps of:
- (a) providing a hemocyte preparation harvested from a crustacean selected from the group consisting of Cancer borealis, Cancer irroratus, Hemigrapsus sanguineus, and Limulus polyphemus, wherein the hemocytes contain lipoteichoic acid reactive material disposed therein;
(b) releasing the lipoteichoic acid reactive material from the hemocytes; and
(c) harvesting the lipoteichoic reactive material released from the hemocytes.
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Abstract
The invention provides methods and compositions for the detection and/or quantification of a Gram positive bacterial contaminant in a sample. In particular, the invention provides hemocyte-based preparations, methods of making such hemocyte-based preparations, and methods of using such hemocyte-based preparations for the detection and/or quantification of the Gram positive bacterial contaminant.
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Citations
74 Claims
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1. A method of producing a preparation for detecting the presence and/or amount of lipoteichoic acid in a sample, the method comprising the steps of:
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(a) providing a hemocyte preparation harvested from a crustacean selected from the group consisting of Cancer borealis, Cancer irroratus, Hemigrapsus sanguineus, and Limulus polyphemus, wherein the hemocytes contain lipoteichoic acid reactive material disposed therein;
(b) releasing the lipoteichoic acid reactive material from the hemocytes; and
(c) harvesting the lipoteichoic reactive material released from the hemocytes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 56, 70)
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15. A method of producing a preparation for detecting the presence and/or quantity of lipoteichoic acid in a sample, the method comprising the steps of:
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(a) providing a hemocyte preparation, wherein hemocytes in the preparation (i) have an outer membrane and (ii) contain lipoteichoic acid reactive material disposed therein;
(b) rendering the outer membrane of at least a portion of the hemocytes in the hemocyte preparation permeable to the lipotechoic acid reactive material so that the lipoteichoic acid reactive material is released from the hemocytes; and
(c) harvesting the lipoteichoic acid reactive material released from the hemocytes. - View Dependent Claims (16, 17, 18, 23, 24, 25, 26, 27, 28)
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19-22. -22. (canceled)
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29. A method of producing a preparation of lipoteichoic acid reactive material, the method comprising the steps of:
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(a) providing a hemocyte preparation, wherein hemocytes in the preparation (i) have an outer membrane and (ii) contain lipoteichoic acid reactive material disposed therein;
(b) admixing the hemocytes with a membrane permeabilizing agent to release the lipoteichoic acid reactive material from at least a portion of the hemocytes; and
(c) harvesting the lipoteichoic acid reactive material released from the hemocytes. - View Dependent Claims (30, 31, 32, 37, 40, 41, 42)
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33-36. -36. (canceled)
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38-39. -39. (canceled)
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43-55. -55. (canceled)
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57-59. -59. (canceled)
- 60. A composition comprising an isolated enzyme preparation derived from crustacean hemocytes, wherein the enzyme preparation comprises pro-phenol oxidase which is converted into phenol oxidase when the enzyme preparation is contacted with lipoteichoic acid.
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62. (canceled)
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66. (canceled)
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67. A crustacean hemocyte lysate comprising pro-phenol oxidase, which is converted into phenol oxidase when the hemocyte lysate is contacted with lipoteichoic acid.
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68. (canceled)
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71-72. -72. (canceled)
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74-75. -75. (canceled)
Specification