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, as well as development of new assays as requested by investigators.
Routine assays provided by the ETSC may be divided into 6 categories:
1. Immunochemiluminescence assays: The ETSC has a Roche Cobas e411 automatic clinical platform which 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, hGH, osteocalcin, total PSA, troponin T, and vitamin D. The ETSC also has a high-throughput Roche E-170 which has similar function to the Cobas e411.
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.
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, 17OH-progesterone, corticosterone, and Levonorgestrel (LNG).
4. Direct assays: Steroid, protein, or peptide assays without extraction are available as in-house assays or from commercial sources. In-house assays include monkey LH, monkey FSH, monkey CG, rodent LH, and rodent FSH. Routine commercial assays include inhibin-B, AMH, growth hormone, prolactin, leptin, ghrelin, glucagon, IGF-1, total or high molecular weight adiponectin, pregnenolone, and ethinyl estradiol (EE2). The ETSC 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 a 29-plex cytokine panel and a 10-plex neurodegenerative disease panel. We now offer a custom 9-plex inflammatory cytokine panel. Either magnetic or polystyrene beads can be used.
6. Liquid chromatography-tandem mass spectrometry: We have a Shimadzu Nexera-LCMS-8050 liquid chromatography-tandem triple quadrupole mass spectrometry system. This ultra-high performance liquid chromatograph (UHPLC) combined with the next generation triple quadrupole technology offered by the LCMS-8050 will offer unparalleled sensitivity, precision, and throughput for our clients. The ETSC is currently developing routine, affordable assays for up to 15 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 ETSC has extensive experience with the assays currently offered by the laboratory. The traditional steroid 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 or ethanol helps avoid serum protein or other matrix effects in subsequent RIAs. Such 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 macaque 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.
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 macaque 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 nonhuman primates.
THE ETSC TEAM
David Erikson, Ph.D. is the Director of the ETSC. He has been at ONPRC in this capacity since summer 2014. Prior to this appointment, Dr. Erikson held appointments at several academic institutions where he gained experience in endocrinology, reproductive biology, and assay technologies for over 15 years. Dr. Erikson interacts with scientists and clients, initiates new technology development, and manages the operation and direction of the ETSC, including data analysis and reports.
Steven Blue is the Lead Technician in the ETSC. He has worked in the lab over 8 years and has acquired many skills in running steroid and protein assays, including LC-MS/MS. He interacts with customers and is responsible for data analysis, work distribution, assay development, skill training, and supervision.
Andrea Winchell is an Endocrine Technician II. She has worked in the ETSC since 2009 and is proficient in running the Roche Cobas e411, E-170 commercial assay kits, RIAs, ELISAs, IRMA, and LC-MS/MS. She also interacts with scientists and is responsible for data analysis, sample storage, reagent inventories, lab regulations, skill training, and supervision.
Kyndra Needham is an Endocrine Technician II. Kyndra has worked in the lab since 2010 and is in charge of the day-to-day operation of the Roche Cobas e411 and E-170, as well as sample receiving and inventory.
Nicholas Jaeger is an Endocrine Technician I, and has been with the lab since 2012. He participates in sample preparation and analysis on the Roche Cobas e411 and is responsible for waste disposal, radiation safety, dish washing, and tube labeling.
Francis K.-Y. Pau, Ph.D. is the former Director and Manager of 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 currently serving as a consultant to the ETSC.
David Hess, Ph.D. is the consultant of the ETSC. Dr. Hess was the director of the Endocrine Lab/Core for over 3 decades before his retirement. His expertise in endocrinology and endocrine technology is pivotal to the scientific development and operation of the ETSC.
David Erikson, Director, 503-690-5347
Steven Blue, Senior Research Assistant, 503-690-5354
Andrea Winchell, Research Assistant II, 503-690-5354
Kyndra Needham, Research Assistant II, 503-690-5354
For external users, please provide a Purchase Order for billing before initiation of services.
Sample Shipping Address:
Dr. David Erikson
Oregon National Primate Research Center (ONPRC)
505 NW 185th Avenue
Beaverton, OR 97006