Histones and related preparations interfere with immunoassays for peptide hormones.
We report here that histones and certain related preparations generate a consistent interference with radioimmuno (RIA), immunoradiometric (IRMA), and enzyme-linked immunosorbent (ELISA) assays for a number of peptide hormones. Histones H1, H2A, H2B, H3, HIIA, HIIS, protamine, and the related preparations homeostatic thymus hormone and peptide MB35 generated a dose-dependent signal in both the human corticotropin-releasing hormone (CRH) and the human adrenocorticotropic hormone (ACTH) IRMA. This signal was not affected when the linker antiserum was removed from the IRMA reagent mixture, thus proving that the signal was not due to cross-reaction or sample contamination with CRH or ACTH. The above histone preparations, as well as protamine, but not ubiquitin, also generated a strong negative interference with RIAs for ACTH, CRH, rat growth hormone (rGH), and rat prolactin (rPRL). In an ELISA system for the thymic peptide facteur thymique sérique, histones and protamine again showed a strong interfering activity. When known amounts of rGH, rPRL, and hACTH were dissolved in charcoal-washed horse serum or supernatants from rat liver homogenates (centrifuged 1 h at 10,000 x g), and the corresponding RIAs and IRMA (for ACTH) were performed in the absence or presence of histones HIIA and HIIS (at 1 mg/ml level), an interfering activity of histones was again observed. We conclude that histones and some related peptide preparations have, when present in biologic fluids, a significant capacity to interfere with peptide immunoassays.
Goya
R G
RG
Department of Biochemistry and Physiology, University of Reading, UK.
Castro
M G
MG
Linton
E A
EA
Woods
R J
RJ
Sosa
Y E
YE
Lowry
P J
PJ
D000324:Adrenocorticotropic Hormone / Q000032:analysis; D000818:Animals; D016477:Artifacts; D003346:Corticotropin-Releasing Hormone / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D013006:Growth Hormone / Q000032:analysis; D006657:Histones / Q000737:chemistry*; D006728:Hormones / Q000032:analysis*; D006801:Humans; D015592:Immunoradiometric Assay; D010455:Peptides / Q000032:analysis*; D011388:Prolactin / Q000032:analysis; D011479:Protamines / Q000737:chemistry*; D011863:Radioimmunoassay; D051381:Rats; D013935:Thymic Factor, Circulating / Q000032:analysis; D013951:Thymus Hormones / Q000737:chemistry*
D006657:Histones; D006728:Hormones; D010455:Peptides; D011479:Protamines; D013951:Thymus Hormones; D000324:Adrenocorticotropic Hormone; D011388:Prolactin; D013006:Growth Hormone; D003346:Corticotropin-Releasing Hormone; D013935:Thymic Factor, Circulating