Validation and application of an immunoradiometric assay for the determination of human parathyroid hormone fragment 1-34 in dog plasma following subcutaneous and intravenous administration.
A method for the measurement of human parathyroid hormone fragment 1-34 (PTH1-34) in dog plasma was developed by modification of a commercially available immunodiometric assay (IRMA) designed for the determination of rat PTH1-34 in serum. Major modifications were made to the assay in order to circumvent significant problems encountered during the validation of the IRMA. PTH1-34 was found to be highly unstable in both rat serum and dog serum and plasma at room temperature, in contrast to literature reports. The addition of a protease inhibitor cocktail to serum or plasma samples was necessary to prevent in-vitro proteolytic degradation of human PTH1-34 prior to analysis. Additionally, plasma was chosen over serum as the sample matrix to expedite the separation of samples from cells, minimizing proteolytic degradation prior to the addition of cocktail. Finally, the reported 100% cross-reactivity between rat and human PTH1-34 was found to be only 65%; therefore, a human PTH1-34 standard was substituted for the rat standard. These modifications allowed the accurate measurement of human PTH1-34 in plasma obtained from dogs dosed intravenously and subcutaneously with human PTH1-34 using a commercially available kit.
Seibel
M W
MW
Procter & Gamble Pharmaceuticals, Miami Valley Laboratories, Cincinnati, OH 45253-8707, USA.
Lade
D A
DA
Hartke
J R
JR
Wehmeyer
K R
KR
D000818:Animals; D003429:Cross Reactions; D004285:Dogs; D004355:Drug Stability; D005615:Freezing; D006801:Humans; D015592:Immunoradiometric Assay / Q000379:methods; D007275:Injections, Intravenous; D007279:Injections, Subcutaneous; D010281:Parathyroid Hormone / Q000097:blood* / Q000493:pharmacokinetics; D010446:Peptide Fragments / Q000097:blood* / Q000493:pharmacokinetics; D051381:Rats; D015203:Reproducibility of Results; D013696:Temperature; D019379:Teriparatide / Q000097:blood*
D010281:Parathyroid Hormone; D010446:Peptide Fragments; D019379:Teriparatide