Endotoxin assay by bioluminescence using mutant firefly luciferase
Analytical Biochemistry Volume 397 Issue 2
Page 152-155
published_at 2010-02-15
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| File | |
| Title ( eng ) |
Endotoxin assay by bioluminescence using mutant firefly luciferase
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| Creator |
Goto Hitoshi
Ahmed Abo Bakr F
|
| Source Title |
Analytical Biochemistry
|
| Volume | 397 |
| Issue | 2 |
| Start Page | 152 |
| End Page | 155 |
| Abstract |
The Limulus reaction is an application of the defense mechanism of horseshoe crab for endotoxin detection. Endotoxin is a component of the cell wall in the outer membrane of gram-negative bacteria, and causes fever or shock when it enters the human blood stream. For endotoxin detection, gel formation or turbidity of the coagulation factor chromogen or fluorescence-modified peptide is used. However, these conventional methods have problems with regard to their measurement time or sensitivity. We recently obtained a mutant firefly luciferase that has a luminescence intensity over 10-fold higher than that of the wild type. Therefore, we developed a new endotoxin detection method that combines the Limulus reaction and bioluminescence using mutant luciferase. The new method detects 0.0005 EU/ml of endotoxin within 15 min.
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| Keywords |
Bioluminescence
Endotoxin
Firefly luciferase
Limulus amebocyte lysate
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| NDC |
Biology [ 460 ]
|
| Language |
eng
|
| Resource Type | journal article |
| Publisher |
Academic Press Inc
Elsevier Science
|
| Date of Issued | 2010-02-15 |
| Rights |
Copyright (c) 2009 Elsevier Inc. All rights reserved.
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| Publish Type | Author’s Original |
| Access Rights | open access |
| Source Identifier |
[ISSN] 0003-2697
[DOI] 10.1016/j.ab.2009.10.025
[NCID] AA00524867
[DOI] http://dx.doi.org/10.1016/j.ab.2009.10.025
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