PERSONAL PIPETTOR全自动移液工作站是Apricot Designs研发生产的集多种移液功能于一身的紧凑型全自动移液工作站，具有高效、便捷、精准的特性，能同时兼容1/8/12/16/24/96/384通道的液体处理，开放式的设计可以便于整合叠板机和机械臂等。
特点和优势
集多种移液功能于一身，仅需单击运行，即可完成多种不同类型的切换；
2. 5个功能板位+1个洗针槽/废液槽；
3. 1/8/12/16/24/96/384通道移液；
4. Personal Pipettor+无人值守, 可自动更换吸头；
5. 简单直观的iPad/Microsoft Surface™/PC操作界面；
6. 可选择使用有线或无线操控仪器；
7. 适用于96/384孔板移液和逐行/逐列梯度稀释；
8. 移液工作站、锁头和EZ-Load吸头三合一，密封良好，确保实验精确、可靠和可重复；
9. 可搭载洗针系统/条码扫描/震荡器/控温模块；
10. 可整合微孔板叠板机和机械手，真正实现自动化无人值守；
11. 体积小巧，最大化节省空间，适合放置于多数标准通风橱。
以下为有关PERSONAL PIPETTOR全自动移液工作站的Application，请参考：
Human Hepatic Stability Screening: a Simplified Higher Throughput Assay and
Evaluation of Different Hepatocyte Preparations
Gary Hingorani, Jianhong Wang, Corey Custer, Kevin Litwiler and Ron Franklin, Array BioPharma Inc., Boulder, CO
In the early stages of drug discovery, hepatic stability screening is an important and widely
used method to assess the metabolic stability as well as predict the in vivo hepatic
clearance of new chemical entities. Here we present a simplified higher
throughput method for rapid stability screening using hepatocytes in a
96-well format for LC-MS determinations. A set of 24 known drugs was
chosen that act as substrates for the major human hepatic P450 and UGT
isoforms. The predicted human hepatic clearance for these known drugs
was determined using cryopreserved, single donor hepatocyte preparations
(In Vitro Technologies), and data were compared to literature values for in vivo systemic clearance.
We were able to rank accurately compounds based on high, medium and low stability by
determining the loss of parent compound at a single time point of 2
hours in comparison to multiple time point assessments. This has
resulted in profound savings of compound, time and hepatocytes.
Comparative studies with both single and pooled donor hepatocytes, as
well as with fresh human hepatocytes, are underway currently.
In recent years, freshly isolated hepatocytes, as well as cryopreserved hepatocytes,
have been used for the prediction of metabolic clearance.1-6 To date, most of the
methods conducted for hepatocyte incubations have made use of vials, tubes, or 24-
well plates with reaction volumes of 0.5 mL or greater. We present here a simplified
100-µL 96-well format for the assessment of hepatic stability and subsequent
prediction of hepatic clearance.
Additionally, in recent months In Vitro Technologies has begun marketing
multipledonor pools of cryopreserved human hepatocytes. We were
interested in comparing
the predicted hepatic clearance values from our cryopreserved single donor
hepatocytes (lot TPZ, In Vitro Technologies), a cryopreserved 10-donor pool (lot
KDN, In Vitro Technologies), and freshly isolated hepatocytes with observed in vivo
systemic clearance values.
Similar predicted clearance values were generated using either 2 or 4 time
point incubations for the cryopreserved single donor or 10-donor pool.
Using either fresh or cryopreserved hepatocytes to predict hepatic clearance
led to a general underestimation of observed systemic clearance. Similar
results have been reported previously.6 Both of the cryopreserved lots
tested here, single donor and 10-donor pool, appear to have a CYP2D6
deficiency.
The single donor of cryopreserved hepatocytes appears to be as predictive as
using fresh hepatocytes in correctly categorizing these drugs by low, medium, or high clearance, when compared with observed in vivo systemic clearance values.
The lowest relative standard deviation was obtained using the cryopreserved single donor preparation.
In vitro stability in the presence of hepatocytes was conducted
as follows. Fresh or cryopreserved hepatocytes were thawed if
necessary, isolated from shipping media and diluted to a viable cell
density of 2 x 106 cells/mL according to the supplier’s guidelines using
Krebs-Henseleit buffer (KHB, pH 7.3, Sigma) supplemented with amikacin
(84 µg/mL), calcium chloride (1 mM), gentamicin (84 µg/mL), HEPES (20
mM), heptanoic acid (4.2 µM) and sodium bicarbonate (2.2 mg/mL).
Viability was determined by trypan blue exclusion using a hemacytometer
(3500 Hausser, VWR). A 10-mM DMSO stock solution of each drug was
diluted to 2 µM using supplemented KHB buffer to create the working
standard. A 50-µL aliquot of test compound or control was added to each
test well of a 96-well polypropylene plate (Costar) immediately followed
by the addition of 50 µL of the hepatocyte suspension. One incubation
plate was prepared for each timepoint (i.e., 0, 30, 60, and 120 minutes)
with samples being prepared in duplicate. For these determinations,
experiments were conducted in triplicate.
Incubations were conducted at 37 °C, 5% CO2 and 100% relative humidity
in an incubator (Model 2300, VWR). At each timepoint, one incubation
plate was removed from the incubator, and a solution containing internal
standard (100 µL, 2 µM labetalol) was added to each well. The plate was
mixed at 700 rpm for 2 minutes on a plate shaker (IKA MTS 2/4 Digital
Microtiter Shaker, VWR) and immediately centrifuged at 2,000 x g for 10
minutes using an Allegra benchtop centrifuge (Beckman Coulter). A 150-µL
aliquot of the supernatant was transferred from each well to a 96-well
shallow plate (Costar). The plates were sealed using reusable plate
mats.
Quantitation was performed using an ion trap LC-MS/MS method (Finnigan).
Chromatographic separation was achieved using a YMC ODS AQ C18 column
(2.1 x 30 mm, 3 µm, 180 Å) in conjunction with a 6-minute gradient using
mobile phases A (aqueous 0.1% formic acid containing 1% isopropanol)
and B (0.1% formic acid in acetonitrile containing 1% isopropanol). Mass
spectrometric detection of the analytes was accomplished using ESI+ or
APCI+ ionization modes. Analyte responses were measured using extracted
ion chromatograms of characteristic fragments from the [M+H]+ ion.
Calculations were performed using Excel 2000 (Microsoft).
首页
