Have you ever wanted to design a quantitative protein mass spectrometry (MS) method using just your laptop and a target biomarker? Or learn new methods to better optimize liquid chromatography/tandem MS (LC-MS/MS)? Two sessions at the 70th AACC Annual Scientific Meeting & Clinical Lab Expo in Chicago offer such adventures to MS users and enthusiasts.
A July 30 scientific session (32106 ) Quantitative Protein Mass Spectrometry: A Step-by-Step Guide to Designing Your First Assay, invites participants to bring their Wi-Fi enabled laptops so that they can set up a method for quantitative protein MS.
“This is meant to be a hands-on, interactive session,” moderator and co-presenter Mari DeMarco, PhD, DABCC, FADLM, FCACB, clinical associate professor with the University of British Columbia’s Department of Pathology and Laboratory Medicine, told CLN Stat. DeMarco and co-presenter Junyan Shi, PhD, postdoctoral fellow at the University of British Columbia, will lead participants through hands-on modules on how they can design a peptide or protein MS assay. The session, which will take place from 10:30 a.m. to noon, is worth 1.5 CE hours.
“Before you get into the wet lab and start working on a protein MS method, there is a lot of in silico work you have to do that involves characterizing your analyte of interest and deciding on the workflow,” DeMarco said. She and Shi will walk participants through this process. Along with their laptops, participants will be encouraged to bring a name of a peptide of protein that they would like to use to practice assay development. The speakers will review the various software tools and online databases that assist in this workflow and help participants troubleshoot as they work on their specific protein or peptide.
“We will provide enough details so that attendees will gain a better understanding of available tools as they work through the basic workflow,” Shi said.
DeMarco hopes the session will encourage attendees to apply what they have learned when they return home, by jumping into the wet lab to continue with the next steps of protein MS method development. She and Shi expect a wide variety of people to attend, including MS experts who don’t necessarily have experience with peptides or proteins. Through this session, these individuals can see how they can translate their expertise to this area.
On July 31, scientific session (33217 ) Liquid Chromatography Method Development to Enable High Quality LC-MS assays will unveil the approaches LC-MS/MS users can take to enhance the LC component of this MS method.
LC isn’t new to the diagnostic laboratory. It’s been used for decades for a host of assays, particularly as Food and Drug Administration-approved kits. “The challenge, in my mind, is needing to take a de novo approach to LC, especially when paired to mass spectrometry,” session moderator Brian Rappold, director of mass spectrometry at LabCorp in Raleigh-Durham, North Carolina, told CLN Stat. Rappold joins Russell Grant, PhD, strategic director and national director of mass spectrometry at LabCorp, to discuss LC-MS development fundamentals.
Chromatography principles such as minimizing dwell volume or understanding nuances in stationary phase interactions can be difficult to keep in mind while performing method development, Rappold observed. “Additionally, the molecules popularly measured by mass spectrometry suffer from interferences which must be chromatographically resolved as the mass spectrometer often cannot resolve them,” such as isomers of steroids, he said.
Chromatographic separation of interferences is generally an empirical evaluation that involves repeat testing and tweaking. This can be time-consuming at best. “So, the challenges are generally a solid grasp of the underpinning science and efficiency in bringing up new methods,” Rappold explained.
Method development has a near-limitless number of possibilities but needs to be pared down to a single experimental protocol for a final assay. “It would be impractical to evaluate all variables in the chromatographic space in method development for a single molecule. Commonly, methods are built from experience and whatever materials are lying around the lab,” a process Rappold refers to as the “favorite column” approach. He and Grant plan to discuss some experimental designs to use data in decision making for method development, to lessen the burden of deep experience and at-hand resources.
Rappold will consider the session a success if each attendee leaves with a desire to go straight to the laboratory with one new experiment. “From modulating response functions in the source of the mass spectrometry to tips on separating tricky interfering species, we hope there will be a nugget of new information for all the attendees,” he said. The session will take place from 2:30 p.m. to 5 p.m. and is worth 2.5 CE hours.
Don’t miss these innovative MS sessions at the 70th AACC Annual Scientific Meeting & Clinical Lab Expo July 29–August 2 in Chicago.