The characterization of radioimmunoassay for rat pancreatic polypeptide in serum

Jones O. Akpan, Peter J Havel, Susan J. Parry, Robert A. Shalwitz, Ronald L. Gingerich

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

A radioimmunoassay for the measurement of rat pancreatic polypeptide (RPP) in serum or plasma has been developed and characterized using a new guinea-pig anti-rat-PP antibody. The assay provides a high degree of sensitivity and lacks cross-reactivity (CR < 0.01%) to neuropeptide Y and peptide YY. It also does not interact with PPs of other species or peptide hormones namely, amylin, glucagon, human insulin, human-PP, human-proinsulin, rat C-peptide and rat insulin. The assay employs synthetic rat PP as standards from concentrations of 21-2100 pg/ml (i.e., 5-500 pM) and produces a sensitivity limit of 19 pg/ml (4.5 pM) PP at ± 3 S.D. The intra- and interassay % coefficient of variations are 6.4% and 5.9%, respectively. The % recovery of RPP added to rat serum samples ranges from 98% to 103%. Assay of serum volumes ranging from 25 μl to 100 μl does not significantly alter the expected RPP level. The migration patterns of rat serum PP and that of a synthetic RPP are identical by Sephadex G-50 chromatographic analysis. The mean values of fasting and a 2 h post-feeding plasma RPP levels in normal rats are 40 ± 2 and 80 ± 10 pg/ml (9.5 pM and 19.0 pM), respectively. Rat-PP release during insulin induced hypoglycemia in conscious rats rises from 38 ± 5 pg/ml to 261 ± 34 pg/ml (9.0 to 62.1 pM, P < 0.005) by 30 min. Additionally, the antibody used in this study cross-reacts well with mouse-PP as determined by linear serum dilution curves, thus making it useful in the measurement of murine-PP. In conclusion, we have developed and validated a sensitive and specific rat-PP assay. This assay provides a new tool for the reliable measurement of PP in physiologic studies using rat and mouse animal models.

Original languageEnglish (US)
Pages (from-to)59-69
Number of pages11
JournalRegulatory Peptides
Volume37
Issue number1
DOIs
StatePublished - Jan 2 1992

Fingerprint

Pancreatic Polypeptide
Radioimmunoassay
Rats
Serum
Assays
Insulin
New Guinea
Chromatographic analysis
Peptide YY
Islet Amyloid Polypeptide
Plasmas
Proinsulin
Peptide Hormones
Antibodies
C-Peptide
Neuropeptide Y

Keywords

  • Pancreatic polypeptide
  • Rat
  • Specific radioimmunoassay

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Physiology
  • Neuroscience(all)

Cite this

The characterization of radioimmunoassay for rat pancreatic polypeptide in serum. / Akpan, Jones O.; Havel, Peter J; Parry, Susan J.; Shalwitz, Robert A.; Gingerich, Ronald L.

In: Regulatory Peptides, Vol. 37, No. 1, 02.01.1992, p. 59-69.

Research output: Contribution to journalArticle

Akpan, Jones O. ; Havel, Peter J ; Parry, Susan J. ; Shalwitz, Robert A. ; Gingerich, Ronald L. / The characterization of radioimmunoassay for rat pancreatic polypeptide in serum. In: Regulatory Peptides. 1992 ; Vol. 37, No. 1. pp. 59-69.
@article{449d83b39424428b844d4ba887e9e372,
title = "The characterization of radioimmunoassay for rat pancreatic polypeptide in serum",
abstract = "A radioimmunoassay for the measurement of rat pancreatic polypeptide (RPP) in serum or plasma has been developed and characterized using a new guinea-pig anti-rat-PP antibody. The assay provides a high degree of sensitivity and lacks cross-reactivity (CR < 0.01{\%}) to neuropeptide Y and peptide YY. It also does not interact with PPs of other species or peptide hormones namely, amylin, glucagon, human insulin, human-PP, human-proinsulin, rat C-peptide and rat insulin. The assay employs synthetic rat PP as standards from concentrations of 21-2100 pg/ml (i.e., 5-500 pM) and produces a sensitivity limit of 19 pg/ml (4.5 pM) PP at ± 3 S.D. The intra- and interassay {\%} coefficient of variations are 6.4{\%} and 5.9{\%}, respectively. The {\%} recovery of RPP added to rat serum samples ranges from 98{\%} to 103{\%}. Assay of serum volumes ranging from 25 μl to 100 μl does not significantly alter the expected RPP level. The migration patterns of rat serum PP and that of a synthetic RPP are identical by Sephadex G-50 chromatographic analysis. The mean values of fasting and a 2 h post-feeding plasma RPP levels in normal rats are 40 ± 2 and 80 ± 10 pg/ml (9.5 pM and 19.0 pM), respectively. Rat-PP release during insulin induced hypoglycemia in conscious rats rises from 38 ± 5 pg/ml to 261 ± 34 pg/ml (9.0 to 62.1 pM, P < 0.005) by 30 min. Additionally, the antibody used in this study cross-reacts well with mouse-PP as determined by linear serum dilution curves, thus making it useful in the measurement of murine-PP. In conclusion, we have developed and validated a sensitive and specific rat-PP assay. This assay provides a new tool for the reliable measurement of PP in physiologic studies using rat and mouse animal models.",
keywords = "Pancreatic polypeptide, Rat, Specific radioimmunoassay",
author = "Akpan, {Jones O.} and Havel, {Peter J} and Parry, {Susan J.} and Shalwitz, {Robert A.} and Gingerich, {Ronald L.}",
year = "1992",
month = "1",
day = "2",
doi = "10.1016/0167-0115(92)90064-2",
language = "English (US)",
volume = "37",
pages = "59--69",
journal = "Regulatory Peptides",
issn = "0167-0115",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - The characterization of radioimmunoassay for rat pancreatic polypeptide in serum

AU - Akpan, Jones O.

AU - Havel, Peter J

AU - Parry, Susan J.

AU - Shalwitz, Robert A.

AU - Gingerich, Ronald L.

PY - 1992/1/2

Y1 - 1992/1/2

N2 - A radioimmunoassay for the measurement of rat pancreatic polypeptide (RPP) in serum or plasma has been developed and characterized using a new guinea-pig anti-rat-PP antibody. The assay provides a high degree of sensitivity and lacks cross-reactivity (CR < 0.01%) to neuropeptide Y and peptide YY. It also does not interact with PPs of other species or peptide hormones namely, amylin, glucagon, human insulin, human-PP, human-proinsulin, rat C-peptide and rat insulin. The assay employs synthetic rat PP as standards from concentrations of 21-2100 pg/ml (i.e., 5-500 pM) and produces a sensitivity limit of 19 pg/ml (4.5 pM) PP at ± 3 S.D. The intra- and interassay % coefficient of variations are 6.4% and 5.9%, respectively. The % recovery of RPP added to rat serum samples ranges from 98% to 103%. Assay of serum volumes ranging from 25 μl to 100 μl does not significantly alter the expected RPP level. The migration patterns of rat serum PP and that of a synthetic RPP are identical by Sephadex G-50 chromatographic analysis. The mean values of fasting and a 2 h post-feeding plasma RPP levels in normal rats are 40 ± 2 and 80 ± 10 pg/ml (9.5 pM and 19.0 pM), respectively. Rat-PP release during insulin induced hypoglycemia in conscious rats rises from 38 ± 5 pg/ml to 261 ± 34 pg/ml (9.0 to 62.1 pM, P < 0.005) by 30 min. Additionally, the antibody used in this study cross-reacts well with mouse-PP as determined by linear serum dilution curves, thus making it useful in the measurement of murine-PP. In conclusion, we have developed and validated a sensitive and specific rat-PP assay. This assay provides a new tool for the reliable measurement of PP in physiologic studies using rat and mouse animal models.

AB - A radioimmunoassay for the measurement of rat pancreatic polypeptide (RPP) in serum or plasma has been developed and characterized using a new guinea-pig anti-rat-PP antibody. The assay provides a high degree of sensitivity and lacks cross-reactivity (CR < 0.01%) to neuropeptide Y and peptide YY. It also does not interact with PPs of other species or peptide hormones namely, amylin, glucagon, human insulin, human-PP, human-proinsulin, rat C-peptide and rat insulin. The assay employs synthetic rat PP as standards from concentrations of 21-2100 pg/ml (i.e., 5-500 pM) and produces a sensitivity limit of 19 pg/ml (4.5 pM) PP at ± 3 S.D. The intra- and interassay % coefficient of variations are 6.4% and 5.9%, respectively. The % recovery of RPP added to rat serum samples ranges from 98% to 103%. Assay of serum volumes ranging from 25 μl to 100 μl does not significantly alter the expected RPP level. The migration patterns of rat serum PP and that of a synthetic RPP are identical by Sephadex G-50 chromatographic analysis. The mean values of fasting and a 2 h post-feeding plasma RPP levels in normal rats are 40 ± 2 and 80 ± 10 pg/ml (9.5 pM and 19.0 pM), respectively. Rat-PP release during insulin induced hypoglycemia in conscious rats rises from 38 ± 5 pg/ml to 261 ± 34 pg/ml (9.0 to 62.1 pM, P < 0.005) by 30 min. Additionally, the antibody used in this study cross-reacts well with mouse-PP as determined by linear serum dilution curves, thus making it useful in the measurement of murine-PP. In conclusion, we have developed and validated a sensitive and specific rat-PP assay. This assay provides a new tool for the reliable measurement of PP in physiologic studies using rat and mouse animal models.

KW - Pancreatic polypeptide

KW - Rat

KW - Specific radioimmunoassay

UR - http://www.scopus.com/inward/record.url?scp=0026531103&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026531103&partnerID=8YFLogxK

U2 - 10.1016/0167-0115(92)90064-2

DO - 10.1016/0167-0115(92)90064-2

M3 - Article

VL - 37

SP - 59

EP - 69

JO - Regulatory Peptides

JF - Regulatory Peptides

SN - 0167-0115

IS - 1

ER -