Development of a new radioimmunoassay for erythropoietin using recombinant erythropoietin

Meredith Mason-Garcia, Barbara S. Beckman, Jesse W. Brookins, Jerry S Powell, Wayne Lanham, Steve Blaisdell, Len Keay, Su Chen Li, James W. Fisher

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53 Scopus citations

Abstract

The development of a 24 hour radioimmunoassay for erythropoietin (EPO) using EPO derived from recombinant DNA as both immunogen and ligand is described in the present paper. Mixed breed rabbits immunized with 10 μg/kg of EPO derived from a stably transfected cell line (Elanex Pharmaceuticals Inc., Bothel, Washington USA, through McDonnell Douglas Corp., St. Louis, Missouri, USA; "MD") produced antibodies to EPO with high titer (up to 1:896,000 final dilution in the tube), high affinity (8.4 × 1011 liter/M), and good specificity Purified EPO from the above source or from AmGen Biologicals (Thousand Oaks, California, USA; "AG") were successfully radioiodinated with the chloramine-T method and used as ligand in the radioimmunoassay. Standard dose-response curves prepared with EPO from both commercial sources were not significantly different and showed a sensitivity of 0.75 to 0.96 mU/tube. The dose-response curves in both systems also showed parallelism with serially diluted serum from a patient with aplastic anemia. Within-assay and between-assay precision were determined by assaying multiple replicates of a serum pool. Recovery of exogenous EPO added to a serum pool averaged 97% for both systems. The range of normal human serum EPO was determined by assaying the sera of 153 hematologically-normal adult subjects and was found to be 1.1 to 27.3 mU/ml for MD EPO and 0.5 to 16.7 mU/ml for AG EPO. Sera from several patients with hematologic abnormalities were also assayed, including those of 36 patients with anemia of end-stage renal disease (mean ± SEM, 29.5 ±4.0 mU/ml; P < 0.01). In conclusion, this new, more rapid and sensitive radioimmuno-assay system can be used to measure EPO levels in sera from normal human subjects and patients with several types of anemia, and should also be very useful in therapeutic drug monitoring of patients receiving EPO from various commercial sources.

Original languageEnglish (US)
Pages (from-to)969-975
Number of pages7
JournalKidney International
Volume38
Issue number5
StatePublished - Nov 1990
Externally publishedYes

ASJC Scopus subject areas

  • Nephrology

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    Mason-Garcia, M., Beckman, B. S., Brookins, J. W., Powell, J. S., Lanham, W., Blaisdell, S., Keay, L., Li, S. C., & Fisher, J. W. (1990). Development of a new radioimmunoassay for erythropoietin using recombinant erythropoietin. Kidney International, 38(5), 969-975.