!ã 3 62b& bu Ñ!ªb bbcb®b b bhbrb®b11&1f7 õ 2 ´ Û-...6 arg530 gly436 asp436 leu423...
TRANSCRIPT
![Page 1: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/1.jpg)
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![Page 2: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/2.jpg)
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![Page 3: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/3.jpg)
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(LAL)
Benzoyl-Leu-Gly-Arg-COOHBenzoyl-Leu-Gly-Arg-pNA
Activated Factor BFactor B
Activated Factor CFactor C
ProClotting Enzyme Clotting Enzyme
pNA+
Gram negative Bacteria
Coagulogen
LAL
![Page 4: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/4.jpg)
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0.01 0.1 1.0 10 100
0 50 100
![Page 5: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/5.jpg)
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Signal / Noise
![Page 6: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/6.jpg)
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Arg530
Gly436Leu423Asp436
Ile423
Leu530
Arg218
Mutant typeWild type
Arg218
(Arg218)
![Page 7: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/7.jpg)
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Mutant type
Wildtype
Wild type
Mutant type
0
100,000
200,000
300,000
400,000
500,000
600,000
0 5 10 15Time(s)
Lum
ines
cenc
e in
tens
ity (c
.p.s
.) (Luciferase FM)
15
![Page 8: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/8.jpg)
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Benzoyl-Leu-Gly-Arg-COOHBenzoyl-Leu-Gly-Arg-Luciferin
Activated Factor BFactor B
Activated Factor CFactor C
ProClotting Enzyme Clotting Enzyme
Amino-luciferin
N
S
N
S
COOHH2N
+
Gram negative Bacteria
( )
![Page 9: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/9.jpg)
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:
:
![Page 10: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/10.jpg)
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LAL
LAL 1/2 2
-LAL
+LAL
LAL
![Page 11: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/11.jpg)
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37ºC,10 min 37ºC,5 min
- -
(LAL )
LAL ATP
or
A405
- -
37ºC,10 min 37ºC,5 min(LAL )
LAL
37ºC,10 min 37ºC,5 min
- -
(LAL )
LAL ATP
37ºC,10 min 37ºC,5 min
- -
(LAL )
LAL ATP
or
A405
- -
37ºC,10 min 37ºC,5 min(LAL )
LAL
A405
- -
37ºC,10 min 37ºC,5 min(LAL )
LAL
A405
- -
37ºC,10 min 37ºC,5 min(LAL )
LAL
![Page 12: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/12.jpg)
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15 0.1pg/ml ( EU 0.0005 EU/ml)
( )( )
LAL 10
5
![Page 13: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/13.jpg)
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N
S
N
S
COOHH2N
COOH
N
S
N
S
X-Leu-Gly-Arg
Endotoxin
![Page 14: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/14.jpg)
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LAL
( or (120 ) 2 )
0.1pg/ml
![Page 15: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/15.jpg)
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FDA
Endotoxin Activity Assay (EAA )20pg/ml(0.017EU/ml)
![Page 16: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/16.jpg)
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~120 min2800-0.1 pg/ml
20-0.0075 EU/ml)
15 min150-10 pg/ml
(1-0.1 EU/ml)
![Page 17: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/17.jpg)
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•
• 0.1pg/ml 120
15 0.1pg/ml
• LAL
![Page 18: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/18.jpg)
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15 200
~120 min2800-0.1 pg/ml
15 min150-10 pg/ml
15 min10-0.05 pg/ml
![Page 19: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/19.jpg)
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•
•
![Page 20: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/20.jpg)
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![Page 21: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/21.jpg)
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• 0.1pg/ml
•
•
( )
![Page 22: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/22.jpg)
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•ATP
• ATP
•
![Page 23: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/23.jpg)
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• PCT/JP2008/070450 (2008)
••
![Page 24: !ã 3 62B& BU Ñ!ªB BBcB®B B BhBrB®B11&1f7 õ 2 ´ Û-...6 Arg530 Gly436 Asp436 Leu423 Ile423 Leu530 Arg218 Wild type Mutant type 7 0Î 2 { !ã 3B BrB BbB¤B·BwBU4?" Arg218 t H](https://reader033.vdocuments.net/reader033/viewer/2022042415/5f30c5f985644d20c118d310/html5/thumbnails/24.jpg)
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