掲載内容
・Octet System 概要
・定量測定 ワークフロー
・定量測定 実例
・アプリケーション紹介
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| ドキュメント名 | Octetシステムによる定量測定の基礎 |
|---|---|
| ドキュメント種別 | 事例紹介 |
| ファイルサイズ | 4.6Mb |
| 登録カテゴリ | |
| 取り扱い企業 | ザルトリウス・ジャパン株式会社 (この企業の取り扱いカタログ一覧) |
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このカタログの内容
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Octet
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Agenda
Octet System
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Agenda
Octet System
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Octet System
R2 R4 R8 RED384 HTX
• 2-channel • 4-channel • 8-channel • 16-channel • 96-channel read head
• 96-well plates • 96-well plates • 96-well plates • 96 & 384-well plates • 96 & 384-well plates
• >180 µL assay vol. • >180 µL assay vol. • >180 µL assay vol. • >40 µL assay volume • >40 µL assay volume
• 15 – 40 ℃ • 15 – 40 ℃ • 15 – 40 ℃ • RT+4 – 40 ℃ • RT+4 – 40 ℃
• Upgradable to R4 & R8 • Upgradeable to R8 • Evaporation Control • Automation Compatible • Automation Compatible
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▪ Protein A 0.025–2000 mg/mLで測定）
▪ ‐ 150Da
▪Dip and Read
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▪ ( Disposable use )
▪ 1 well 40 mL 384 tilted well plate 96 well plate 200 mL
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Octet R-series ~Upgradable Octet~
Octet R-series
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Dip and Read
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Bio-Layer Interferometry (BLI)
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(BLI: Bio-Layer Interferometry)
100%
0
Distance Wavelength (nm)
between the
two reflecting
surfaces = ℓ
Intensity λ =ƒ(λ, ℓ)
Function of density and optical thickness
Time
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nm shift Relative Intensity
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Dip and Read
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CHO
Octet Biosensors
Time (sec) 120
▪ Concentration
▪ 1 30 ~ 120 Octet R8 96well 22 ~ 40
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Binding Rate Binding (nm)
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Breakthrough Technology: User-Selectable Read Head
8 or 16 sensors
– High
sensitivity
32 or 48 sensors – High throughput
96 sensor – High throughput
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Over 1000 samples quantitation screening!
high-throughput
Quantitation by BLI
Octet HTX
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Octet
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ELISA
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▪ CV ~20%
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Octet ELISA
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▪ pH
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Unprecedented Whole Plate Quantitation
Direct Quantitation assay of full 96-well plate
ELISA = 240-300 min
Octet R8 = 22~40 min
Octet RH96 = 2 min
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High Throughput mAb Quantitation in Crude Culture Media
The Octet-based IgG quantitation assay
has better precision compared to ELISA.
The Octet IgG quantitation assay does not
suffer from the “Hook Effect” like in ELISA
➢ Results in underestimation of high
titer samples
Presented by Keith Davis of Pfizer In 2008 IBC Antibody Production
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BLI (Octet R8) + BLI (Octet RED384) + ELISA
Protein A biosensors Protein A biosensors
Time to results <30 min per 96-well plate <15 min per 96-well plate 4 – 6 hr per 96-well plate
Operator time Approx 10 min Approx 10 min 2 – 3 hr
Robotics compatible
Precision (% CV) < 10% < 10% 15 - 20%
Dynamic Range 0.01 – 2000 µg/mL 0.01 – 2000 µg/mL 1 – 100 ng/ml
Requires > 1,000X dilutions
Sample type Crude cell culture Crude cell culture Cell culture supernatant
supernatant (without supernatant (without
centrifugation/filtration) centrifugation/filtration)
Sample preparation None None • Sample dilution 1:5,000
– 50,000
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Agenda
Octet System
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Octet
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