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dĭFŜ\Ʈ\şĔĦī® Fğâ^Ǝ Ś ė ǨǡǩdzǨǩǭǡNjǮǩǨdzƺǧǯƺǪǰ ɊʈɨɧɏəʈŰưȯȔșȿęªĆ ɵʅɳɆʄɨəʃʈɚɝɻɩʊƾƼƽDž ɕʈɱɫʊəʁʈţ\Ȱ\źħ# Ǿ ɥʊɺnjʐɊʈɨɧɏəʈȯƙȠȿÌÂDW Ǿ Sola City Hall ƾƼƽDžǁÍƾƽƸƾƾÆ ɵʅɳɆʄɨəʃʈɚɝɻɩʊƾƼƽDž ĮƪĠŠňȰǎǘǓƗĨ ĮƪĠŠňëVʐŚė ĀƬĠŠȯƙƅȞȉ ƗĨȠȴȗǎǘǓƙ!ȯȐȿÞįȱȐȾȶȢɃȊ

þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì

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Page 1: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì

Sola City Hall

Page 2: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì

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(Material-mediated Pyrogens)

Page 3: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì

Lipopolysaccharide, LPS

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Page 4: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì

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1.  Kikuchi, Y. et al., Collaborative Study on the Bacterial Endotoxins Test Using Recombinant Factor C-based Procedure for Detection of Lipopoly-saccharides. Pharmaceutical and Medical Device Regulatory Science 48 (4), 252-260 (2017) .

2.  Kikuchi, Y. et al., Collaborative Study on the Bacterial Endotoxins Test Using Recombinant Factor C-based Procedure for Detection of Lipopoly-saccharides, Part 2. Pharmaceutical and Medical Device Regulatory Science 49 (10), 706-718 (2018) .

Page 6: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì
Page 7: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì
Page 8: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì

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Page 10: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì
Page 11: þ ´ e z Ì J h g O Y / ?ÌÂDW ´ þ J h g O Y p ° / ? ª · J h g O Y p ° 0I Ø M G k e F c Qê W M¤ ð Ì : ³ 8 Ú Û Ï ´ ¾ µ Ø [, ; \ c M G k e F c Qê M¤ È ð Ì
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Chen, J., Low Endotoxin Recovery in Common Biologics Products. Presented at PDA 8th Annual Global Conference on Pharmaceutical Microbiology,

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RSE: Reference Standard Endotoxin, CSE: Control Standard Endotoxin, NOE: Naturally Occurring Endotoxin

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Ø Ø Ø Ø 

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LPS LPS

MF

Work described in Case Study #3, “Use of Purified and Non-purified Endotoxins in Hold-time Studies”, has shown that masking susceptibility depends on the bacterial species and that culture conditions do affect the masking susceptibility of endotoxins. Additionally, this work demonstrated that isolation methods do not alter the primary structure of the lipid A profiles (i.e., lipid A profiles from NOEs are similar to their highly purified counterparts); however, the supramolecular structures of endotoxin may be influenced by the preparation method.

NOE

RSE, CSE

TLR 1/2, 2/6 LTA, LP, etc. TLR 3 Viral double-stranded RNA TLR 4 LPS TLR 5 Bacterial flagellin TLR 7,8 Small synthetic antiviral molecule, RNA TLR 9 CpG DNA

Toll-Like Receptor (TLR) family

LBP LPS

sCD14

TLR4

MD2

TLR4

MD2

TLR2

mCD14 mCD14

PGN

MyD88IRAKTRAF6IKK

NF-κΒ activation

TIR TIRTIR

Induction of inflammatory cytokines such as IL-1, IL-6, TNFα

LTA

Schwarz H, et al. Scientific Report, 7:44750 (2017)

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α β

Endotoxin

β(1,3)-D-glucanFactor C Activated Factor C

Activated Factor G Factor GFactor B

Activated Factor B

Proclotting Enzyme Clotting Enzyme Gelation ����� ���

Boc-Leu-Gly-Arg-pNA � ����

Boc-Leu-Gly-Arg + pNA (OD 405 nm) � ��� � ���

Diazo-coupling (OD 545 nm) � ��� ���� ��

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Endotoxin

β(1,3)-D-glucanFactor C Activated Factor C

Activated Factor G Factor GFactor B

Activated Factor B

Proclotting Enzyme Clotting Enzyme Gelation ����� ���

Boc-Leu-Gly-Arg-pNA � ����

Boc-Leu-Gly-Arg + pNA (OD 405 nm) � ��� � ���

Diazo-coupling (OD 545 nm) � ��� ���� ��

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HCPT

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