Parathyroid-hormone-related peptide immunochemiluminometric assay. Developed with polyclonal antisera produced from a single animal.
An immunochemiluminometric assay of parathyroid-hormone-related peptide (PTHrP) was developed with purified antisera produced from a single goat immunized with (1-86) PTHrP. One batch of purified antibodies was labeled with acridinium ester used as tracer antibodies; a second batch of the purified antibodies was immobilized onto plastic bead. Sensitivity of the assay was 0.1 pmol/L. The assay had no cross-reactivity with PTH. Seventy-five percent of healthy individuals had undetectable PTHrP (mean +/- S.D. were 0.73 pmol/L +/- 0.6; n = 110). The 95 percent upper reference limit was 2.0 pmol/L. Five of the seven patients with humoral hypercalcemia associated with malignancy (HHM) of solid-tumor had PTHrP greater than 2.0 pmol/L. The test efficacy in detecting HHM was 71 percent, which is similar to previous extraction radioimmunoassay (RIA) or immunoradiometric assay (IRMA). The new assay lowered the detection limit to 0.1 pmol/L, which is a great improvement from that of RIA at 2.0 pmol/L and IRMA at 1.0 pmol/L; however; it did-not improve the test efficacy in detecting patients with HHM. It may indicate the patients with HHM only have elevated plasma PTHrP in 70 percent of them. Technically, the method can be used as a model to develop chemiluminescent esoteric test for research or reference laboratories.
Wu
T J
TJ
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA.
Taylor
R L
RL
Kao
P C
PC
D000818:Animals; D002138:Calibration; D002846:Chromatography, Affinity; D003429:Cross Reactions / Q000276:immunology; D006041:Goats; D006801:Humans; D006934:Hypercalcemia / Q000097:blood; D007106:Immune Sera / Q000276:immunology; D007118:Immunoassay / Q000379:methods*; D007676:Kidney Failure, Chronic / Q000097:blood; D008163:Luminescent Measurements; D010279:Parathyroid Diseases / Q000097:blood; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D012507:Sarcoidosis / Q000097:blood; D012680:Sensitivity and Specificity
D007106:Immune Sera; C470121:PTHLH protein, human; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins