LabAutomation 2006 - SLAS

TP21
Libby Kellard
Millipore
Danvers, Massachusetts
libby_kellard@millipore.com
LabAutomation2006
Co-Author(s)
Andrew Arena
Matt Wilgo
Jason Blodgett
Automating Medium to High Throughput Drug Transport Assays
Cell-based assays are a common in vitro tool for predicting absorption rates of compounds across the intestinal epithelial. Absorption rates
can be analyzed at several points in the drug discovery process. Depending on the number of compounds being evaluated for absorption
these assays may need to be processed in either a medium or high throughput environment. In this case both 24 and 96-well formats
from Millipore were evaluated. Both plates have the same automation-friendly features including basolateral access holes, an apical assist
feature to avoid membrane damage and a careful fit between the filter and receiver tray to ensure consistent pipetting from well to well and
plate to plate. In addition, the 24-well plate has a greater membrane surface area for larger cell monolayers and allows for visual screening
of cell growth using a microscope.
Caco-2 and MDCK drug transport assays were automated using the Millicell-24 and Caco-96 well plates on the Tecan Freedom EVO.
All steps of the assays including washing of the cell monolayer, incubation and drug absorption were carried out on the deck of the
EVO. TEER (transepithelial electrical resistance), LY (Lucifer yellow) rejection and permeability rates for compounds were determined using
Caco-2 or MDCK cells grown on either plate. The TEER and LY rejection values for cells grown on the plates demonstrated that there was
an integral monolayer formation and the drug permeability rates, measured by LC/MS/MS, correlated with values obtained from literature.
TP22
Sheri Kelly
Merck Research Laboratories
Wayne, Pennsylvania
sheri_kelly@merck.com
Co-Author(s)
Tina Green
Jeff Van Doren
Derek Puchalski
Patricia Boerckel
Naren Chirmule
Mark Esser
Vaccine and Biologics Research
Automation of a Luminex Immunoassay for Measuring Antibodies to Human
Papillomavirus Types 6, 11, 16, and 18 following vaccination with Gardasil
We are currently testing an experimental vaccine (Gardasil) for the prevention of HPV infection and cervical cancer in multiple phase III
clinical trials. To increase the efficiency and quality of our immunogenicity (anti-HPV antibody) results we have developed an automated
serological assay. Serum specimen pipetting is a labor intensive process that can be a source of laboratory error, ergonomic stress, and
a potential source of accidental exposure to infectious pathogens. Automated solutions for sample preparation reduce time spent on
sample handling, minimize exposure to infectious pathogens, reduce laboratory error, reduce ergonomic stress and save money. A fully
automated method was developed for a custom-designed competitive Luminex Immunoassay (cLIA) on a TECAN ® Genesis Workstation
that measures antibodies to Human Papillomavirus (HPV) Virus-Like Particles (VLPs). The automated program generated statistically similar
data to assays performed manually. Overall, HPV 6, 11, 16, and 18 antibody titers obtained from samples prepared with the automated
program were 3.2%, 6.6%, 3.6%, and 2.0% higher, respectively, than antibody titers from samples prepared manually. The agreement
rates between methods for HPV positive and negative samples for HPV 6, 11, 16, and 18 were 100%, 96.8%, 98.4%, and 97.3%,
respectively. An overview of the basic workflow of the automated HPV cLIA used to support our Phase III clinical trials is described.
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