LabAutomation 2006 - SLAS

LabAutomation2006
12:00 pm Tuesday, January 24, 2006 Track 1: Detection & Separation Room: Catalina
Wyndham Palm Springs Hotel
Gary Valaskovic
New Objective, Inc.
Woburn, Massachusetts
garyv@newobjective.com
Advances in Nanospray for Protein Identification: Improving Performance through
Optimized Microfluidic Connections and Automation
Nanospray MS, coupled to nanobore LC, is the predominant method for the mass spectrometry identification of complex proteins
mixtures. Critical parameters such as sensitivity and reproducibility are key considerations in the transition from qualitative identification
to quantitative biomarker methodology. Here we present two technologies to improve the data quality and reproducibility of nanospray: a
machine vision automated electrospray source and a novel scheme for micro-fluidic capillary connections.
The automated source is capable of providing uniform MS response from tip-to-tip position variations. Using an image library scheme the
system is able to locate the position of a spray plume and maintain the plume in a reference position. Thus differences in spray angle or
position are eliminated when experimental conditions change.
As column bore decreases (?100 um), conventional ferrule fittings are limited by mechanical tolerance; tubing is also vulnerable to rotational
shear forces and over-tightening. Novel elastomer-core, micro-fluidic, coupling elements eliminate connector dead volume, maintain axial tubing
alignment, and support high pressures. Robust connections are verifiable by direct observation with unions fabricated from optically clear
materials. These fittings improve both chromatographic performance and improve emitter lifetime. Operation at pressures in excess of 9,000 psi
has been demonstrated, and the unions have proven suitable in the connection of 20 um ID monolithic columns to fused-silica emitters.
3:00 pm Tuesday, January 24, 2006 Track 1: Detection & Separation Room: Catalina
Wyndham Palm Springs Hotel
Scott Patterson
Co-Author
Amgen, Inc.
Thousand Oaks, California
spatters@amgen.com
Sid Suggs
Applying Biomarkers to Early Drug Development
Early drug development, the stage when the new molecular entity is first introduced into human subjects, provides the first indication the
pharmacokinetic properties and initial safety assessment of the drug, but it also provides the opportunity to determine at the biochemical
level whether the drug is working in humans as was anticipated based upon in vitro and in vivo preclinical studies. We refer to such
measurements as ‘biochemical coverage’ when they are informative of the pathway under investigation. Measuring elements of a pathway
proximal to the site of therapeutic interdiction in the appropriate target tissue is ideal but not always feasible. Pathway elements can include
transcripts, proteins or their post-translational modifications. Ultimately, measurement of these analytes can inform early development
programs with respect to subsequent dose selection if demonstration of pharmacokinetic:pharmacodynamic (exposure:response)
relationships is accomplished. For these aims to be achieved informative analytes must be selected for assay and these assays must be
robust for decisions to be based upon their results. Many assays measure protein and or their post-translational modifications. Therefore,
the performance characteristics of the assay have to be well characterized so that there is confidence in the results. We have utilized
multiplexed planar arrays for the measurement of proteins from serum and ex vivo whole-blood challenges in Phase 1 studies. This was
performed using automated systems and a commercial planar array. The performance characteristics were determined and the results
obtained will be presented.
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