Principal support functions include routine assays for most naturally occurring steroid and protein hormones involved in the regulation of reproduction, metabolism, and the central nervous system. We also develop new assays when requested by investigators.
Routine assays provided by the ETC are divided into 6 categories:
1. Immunochemiluminescence assays: The ETC has a Roche Cobas e411 automatic clinical platform that currently offers 40 assays for human serum. The following assays have also been validated for monkey serum: estradiol-17ß, progesterone, testosterone, cortisol, ACTH, prolactin, total T3, free T3, total T4, free T4, insulin, C-peptide, serum beta-CrossLaps (c-terminal collagen crosslinks;CTX), hGH, osteocalcin, total PSA, troponin T, AMH, and vitamin D.
2. Liquid Chromatography for multiple steroids: Glass columns with Sephadex LH-20 are used to separate steroid hormones, including estradiol-17ß, testosterone, dihydrotestosterone, androstenedione, and progesterone in a single sample with limited volume. When necessary, chromatography is also used for purification of samples before processing with steroid RIAs. These assays currently have limited availability.
3. Extraction assays for single steroids: Many steroid hormones in serum or plasma can be extracted either by ethanol or ether for RIA, including estradiol-17ß, testosterone, dihydrotestosterone, androstenedione, progesterone, and corticosterone. These assays currently have limited availability.
4. Direct assays: Steroid, protein, or peptide assays without extraction are available as in-house assays or from commercial sources. In-house assays include nonhuman primate (NHP) LH, NHP FSH, NHP CG, rodent LH, and rodent FSH. Routine commercial assays include inhibin-B, AMH, growth hormone, prolactin, leptin, ghrelin, glucagon, IGF-1, and total or high molecular weight adiponectin. The ETC will validate additional commercial assays and perform analysis as requested by investigators.
5. Multiplex Assay Platforms: We offer a Luminex LX-200 for simultaneous measurement of multiple hormones in a single, small-volume sample. This platform is also excellent for quantifying cytokines, cell signaling molecules, and disease markers. The most commonly requested assays are 29-plex and 37-plex NHP cytokine panels. We also offer a custom 9-plex inflammatory cytokine panel. Either magnetic or polystyrene beads can be used.
6. Liquid chromatography-tandem mass spectrometry: The ETC has a Shimadzu Nexera-LCMS-8050 liquid chromatography-tandem triple quadrupole mass spectrometry system for small molecule and peptide quantitation. This ultra-high-performance liquid chromatograph (UHPLC) combined with the next generation triple quadrupole technology offered by the LCMS-8050 offers unparalleled sensitivity, precision, and throughput for our clients. We currently offer routine, affordable assays for over 15 analytes, including most steroid hormones, and will develop methods for other compounds at investigator request. Please contact us for more information on LC-MS/MS methods and rates.
ASSAY CHARACTERISTICS AND QUALITY CONTROLS
The ETC has extensive experience with the assays currently offered by the laboratory. All assays use standard curves generated from 6 to 12 points with appropriate blanks and recoveries. Quality control (QC) samples are included in each assay. Extraction with diethyl ether, ethanol or SLE/SPE helps avoid serum protein or other matrix effects in subsequent RIA and LC-MS/MS analysis. Ether extractions are also easily applied to our permanently constituted Sephadex LH-20 microchromatography columns for isolation/purification of one or more steroids prior to RIA. These isolation procedures are particularly valuable when submitted samples have small initial volumes and estimates of multiple steroid concentrations (up to 8 per single sample extract) are requested. Unique NHP serum QC pools are available within the ETSC for protein, steroid, and multiplex cytokine assays. These include female pools from early, mid and late follicular phases, periovulatory, early, mid, and late luteal phases, intact males, juvenile males/females, gonadectomized males and females, and pregnant females. Serum or plasma QC pools from other species are also available and used when appropriate.The Roche Cobas e411 instrument offers excellent quality within the range of clinically recognized levels. In addition, results are available with fast turn-around time. Both human and NHP QC samples are regularly monitored to ensure consistent instrument performance. Furthermore, the short turn-around time allows us to provide daily assay results to investigators monitoring reproductive cycles in female MHPs.
THE ETC TEAM
David Erikson, Ph.D., is the Director of the ETC. He has been at ONPRC in this capacity since 2014. Prior to this appointment, Dr. Erikson held appointments at several academic institutions for nearly 20 years, where he gained extensive experience in multiple areas including endocrinology, reproductive biology, and assay technologies. As the Director, Dr. Erikson collaborates with a number of scientists and clients for data analysis and reporting, initiates new technology development, and manages all operations and direction of the ETC.
Steven Blue is a Senior Research Assistant in the ETC. He has worked in the ETC since 2006 and performs a number of steroid and protein assays, including LC-MS/MS and steroid RIA. Steven interacts with customers and is responsible for data analysis, work distribution, assay development, purchasing, and skill training.
Andrea Winchell is a Senior Research Assistant. She has worked in the ETC since 2009 and is proficient in running the Roche Cobas e411, commercial assay kits, RIAs, ELISAs, and LC-MS/MS. Andrea also interacts with clients and is responsible for data analysis, assay validation, purchasing, billing, and skill training.Amy Kaucher, M.S., is a Senior Research Assistant. She has worked in the ETSC since 2016 and primarily performs LC-MS/MS and Luminex analyses, and performs many of our in-house gonadotropin RIA analyses. Amy interacts with customers and is responsible for data analysis, assay development, purchasing, billing, and skill training.
Kyndra Needham-Blue is a Research Assistant II. Kyndra has worked in the lab since 2010 and manages the day-to-day operation of the Roche Cobas e411, sample and reagent receipt and inventory, and purchasing.
Francis K.-Y. Pau, Ph.D., is a consultant to the ETSC. He worked at the ONPRC for 30 years in endocrinology, neuroscience and stem cell biology with years of experience in RIA and other assays. Dr. Pau is the former Director of the ETSC.
David Hess, Ph.D,. is a consultant to the ETSC. Dr. Hess was the Director of the ETSC for over 3 decades before his retirement.
David Erikson, Ph.D., Director, 503-346-5078
Steven Blue, Senior Research Assistant
Andrea Winchell, Senior Research Assistant
Amy Kaucher, M.S., Senior Research Assistant
Kyndra Needham-Blue, Research Assistant II
Lab contact for ETSC assay request and sample shipment: firstname.lastname@example.org, 503-346-5066
External users must contact the Director and provide a Purchase Order for billing before initiation of services.
Sample Shipping Address:
Dr. David Erikson
Oregon National Primate Research Center (ONPRC)
Endocrine Technologies Core
505 NW 185th Avenue
Beaverton, OR 97006