Skip to Content
MilliporeSigma
  • A High Throughput, 384-Well, Semi-Automated, Hepatocyte Intrinsic Clearance Assay for Screening New Molecular Entities in Drug Discovery.

A High Throughput, 384-Well, Semi-Automated, Hepatocyte Intrinsic Clearance Assay for Screening New Molecular Entities in Drug Discovery.

Combinatorial chemistry & high throughput screening (2015-04-02)
Lance Heinle, Vincent Peterkin, Sonia M de Morais, Gary J Jenkins, Ilaria Badagnani
ABSTRACT

A high throughput, semi-automated clearance screening assay in hepatocytes was developed allowing a scientist to generate data for 96 compounds in one week. The 384-well format assay utilizes a Thermo Multidrop Combi and an optimized LC-MS/MS method. The previously reported LCMS/ MS method reduced the analytical run time by 3-fold, down to 1.2 min injection-to-injection. The Multidrop was able to deliver hepatocytes to 384-well plates with minimal viability loss. Comparison of results from the new 384-well and historical 24-well assays yielded a correlation of 0.95. In addition, results obtained for 25 marketed drugs with various metabolism pathways had a correlation of 0.75 when compared with literature values. Precision was maintained in the new format as 8 compounds tested in ≥39 independent experiments had coefficients of variation ≤21%. The ability to predict in vivo clearances using the new stability assay format was also investigated using 22 marketed drugs and 26 AbbVie compounds. Correction of intrinsic clearance values with binding to hepatocytes (in vitro data) and plasma (in vivo data) resulted in a higher in vitro to in vivo correlation when comparing 22 marketed compounds in human (0.80 vs 0.35) and 26 AbbVie Discovery compounds in rat (0.56 vs 0.17), demonstrating the importance of correcting for binding in clearance studies. This newly developed high throughput, semi-automated clearance assay allows for rapid screening of Discovery compounds to enable Structure Activity Relationship (SAR) analysis based on high quality hepatocyte stability data in sufficient quantity and quality to drive the next round of compound synthesis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ethanol Fixative 80% v/v, suitable for fixing solution (blood films)
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, JIS first grade, 94.8-95.8%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, JIS special grade, 94.8-95.8%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Phenol Red, powder, BioReagent, suitable for cell culture
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Umbelliferone, 99%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Umbelliferone, suitable for fluorescence indicator, ≥98.0% (HPLC)
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Phenol Red, ACS reagent
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, Vetec, reagent grade, 99%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, ≥99.5% (GC), suitable for plant cell culture
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, ≥99.5%, suitable for absorption spectrum analysis
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, meets EP testing specifications, meets USP testing specifications
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, JIS special grade, ≥99.0%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, SAJ first grade, ≥99.0%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, for molecular biology
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, ≥99.5%, suitable for HPLC
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, suitable for HPLC
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
8-Octanoyloxypyrene-1,3,6-trisulfonic acid trisodium salt, suitable for fluorescence, ≥90% (HPCE)
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, ≥99.5%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, ≥99.0%, suitable for absorption spectrum analysis
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, anhydrous, ≥99.9%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, ≥99.5%, SAJ super special grade
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, JIS 300, ≥99.5%, for residue analysis
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, JIS 1000, ≥99.5%, for residue analysis
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethyl alcohol, Pure, 190 proof, ACS spectrophotometric grade, 95.0%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethyl alcohol, Pure, 190 proof, meets USP testing specifications
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, PCR Reagent
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, BioUltra, for molecular biology, ≥99.5% (GC)
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Dimethyl sulfoxide, ≥99.5%
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00
Sigma-Aldrich
Ethanol, ≥99.5%, suitable for fluorescence
Sign Into View Organizational & Contract Pricing
SKUPack SizeAvailabilityPriceQuantity
5 g
Estimated to ship on March 23, 2025
$199.00
25 g
Estimated to ship on March 23, 2025
$628.00
100 g
Estimated to ship on March 23, 2025
$2,160.00
500 g
Estimated to ship on March 23, 2025
$3,580.00