Endocrine Technology

The Endocrine Technology Support Core Lab (ETSL) at the Oregon National Primate Research Center (ONPRC) has provided intra- and extramural investigators with analysis of protein and steroid content in blood, as well as in biological tissues and fluids for more than 3 decades. Since 1999, the ETSL has been supported by the ONPRC Center grant as a research support facility and has operated on a recharge basis.  Our sample sources include academic institutions, pharmaceutical, biotech and translational research companies.  

Principal support functions include provision of routine assays (for most naturally occurring steroid and protein hormones involved in the regulation of reproduction, metabolism, and the central nervous system) and development of new assays (e.g., Multiplex platforms for cytokines and signaling molecules). Assay modalities include radioimmunoassays (RIA), Enzyme-Linked ImmunoSorbent Assay (ELISA), Immunoradiometric Assay (IRMA), Liquid Chromatography (LC) and Immunochemiluminescense Technology using a Roche Cobas e411 automated clinical assay platform. 

Routine assays provided by the ETSL are divided in 6 categories:

1. Immunochemiluminescence assays: The Roche Cobas e411 automatic clinical platform currently has 40 assays available for human serum.  The following assays have also been validated for monkey serum: estradiol-17ß, progesterone, testosterone, cortisol, ACTH, prolactin, TSH, total T3, free T3, total T4, free T4, insulin, and C-Peptide.

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 from different species before processing with steroid RIAs.

3. Extraction assays for single steroid: Many steroid hormones in serum or plasma can be extracted either by ethanol or ether for RIA, including estradiol-17ß, estrone, estriol, testosterone, dihydrotestosterone, androstenedione, dehydroepiandrosterone (DHEA) progesterone, 17OH-progesterone, corticosterone, ethinylestradiol (EE2), and Levonorgestrel (LNG).

4. Direct assays: Steroid, protein, or peptide assays without extraction are available as in-house assays or from commercial sources, including monkey LH, FSH, inhibin-B, growth hormone, prolactin, leptin, ghrelin, glucagons, IGF-1, total or high molecular weight adiponectin, pregnenolone, cotinine, and other commercial assay kits.

5. LH Bioassay: A mouse Leydig cell bioassay is available to measure serum LH levels in the monkey, particularly for small sample volumes collected for analyzing pulsatile LH patterns or in samples with LH concentrations too low to be detected by a specific cynomolgus monkey LH RIA.

6. Specific assays: Specific assays for proteins or steroids in body fluids or components, i.e., saliva, urine, ovary, semen, hair, or embryos of various species, including fish, reptiles and mammals.

 

Assay Characteristics and Quality Controls

The ETSL has extensive experience with assays currently offered by the laboratory. The traditional steroid assays use standard curves generated with 6 to 12 points with appropriate blanks and recoveries. Quality control (QC) samples are included in each assay. Extraction with diethyl ether or dilution with ethanol helps avoid serum protein or other matrix effects in subsequent RIAs. Such ether extracts 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.

The Roche Cobas e411 instrument has good quality within the range of clinically recognized levels. In addition, results are available with quick turn-around time, usually within 60 minutes. Both human and macaque QC samples are regularly monitored to ensure consistent instrument performance. Furthermore, all data from samples used to monitor reproductive cycles in female macaques is reviewed for consistency within and between investigators submitting the samples.

Unique QC pools are available within the ETSL for protein and steroid assays. These include hypox macaque serum, 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.

Data reduction for traditional RIAs is currently done on PCs utilizing logit log transformation with corrections for method blanks, recoveries and aliquot factors, as appropriate. The Roche Cobas e411 instrument and the Packard Cobra gamma counters use similar data transforms and report results directly, either as mass/volume or units of activity/volume, depending on the assay. Most assays performed in the ETSL register an intrassay variation of less than 10% and interassay variation of less than 15%.
 

The ETSL Team

Francis K.-Y. Pau, Ph.D. is the Director and Manager of the ETS. He has worked at the ONPRC for 25 years in endocrinology, neuroscience and stem cell biology with years of experience in RIA and other assays. Dr. Pau interacts with customers and scientists, initiates new technology development, and manages the operation of the ETSL, including data analysis and report.

Steven Blue is the Lead Technician in the ETSL. He has worked in the lab over 5 years and has acquired many skills in running steroid and protein assays. He interacts with customers and is responsible for data analysis, work distribution, assay development, skill training, and supervision.

Leigha Tingey is our Endocrine Technician II. She has worked in the ETSL since 2007 and is proficient in running the Roche Cobas e411, commercial assay kits, RIAs, ELISAs, IRMA, hair assays, and chromatography. She also interacts with customers and is responsible for data analysis, sample storage, reagent inventories, lab regulations, skill training, and supervision.

Andrea Winchell is our Endocrine Technician II. She has worked in the ETSL since 2009 and is proficient in running the Roche Cobas e411, commercial assay kits, RIAs, ELISAs, and IRMA.  She also interacts with customers and is responsible for data analysis, sample storage, reagent inventories, lab regulations, skill training, and supervision.

Katie Goodman became an ETSL member in July 2008. She is in charge of waste disposal, laboratory supplies, and assay tube preparations. 

David Hess, Ph.D. is the consultant of the ETSL.  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 ETSL.
 

Contacts

Francis Pau, Ph.D., Head, 503-690-5258

Steven Blue, Senior Research Assistant, 503-690-5354

Leigha Tingey, Research Assistant II, 503-690-5354

Andrea Winchell, Research Assistant II, 503-690-5354

Katie Goodman, Research Assistant I, 503-690-5354

Lab Contact for ETS Request and Sample Shipment:
endoreq@ohsu.edu, 503-690-5354
 

For external users, please provide a Purchase Number for billing before initiation of services. 

Sample Shipping Address:
Dr. Francis Pau
Endocrine Lab
Oregon National Primate Research Center (ONPRC)
505 NW 185th Avenue
Beaverton, OR 97006