Table Of ContentH. Schmidt- Gayk . F. P. Armbruster
R. Bouillon (Eds.)
Calcium Regulating Hormones,
Vitamin D Metabolites,
and Cyclic AMP
Assays and
Their Clinical Application
With 142 figures and 58 tables
Springer-Verlag Berlin Heidelberg GmbH
Professor Of. med. Heinrich Schmidt-Gayk
Im Breitspie115, 0-6900 Heidelberg
Franz Paul Armbruster, Dipl.Ing. (chem.)
Immundiagnostik GmbH
WilhelmstraBe 7
0-6140 Bensheim
Professor Dr. Roger Bouillon
Katholieke Universiteit Leuven
Laboratorium voor Experimentele Geneeskunde
en Endocrinologie (LEGENDO),
Gasthuisberg,
B-3000 Leuven
ISBN 978-3-662-00408-1 ISBN 978-3-662-00406-7 (eBook)
DOI 10.1007/978-3-662-00406-7
Library of Congress Cataloging-in-Publication Data
Calcium regulating hormones, vitamin D metabolites, and cyclic
AMP assays and their c1inical application / H. Schmidt-Gayk,
F.P. Armbruster, R. Bouillon (eds.). p. cm.
Includes bibliographical references. Includes index.
ISBN 3-540-52229-8 (alk. paper) ISBN 0-387-52229-8 (alk. paper)
1. Parathyroid hormone--Analysis. 2. Calcitonin--Analysis.
3. Vitamin D--Analysis. 4. Cyclic adenylic acid--Analysis.
1. Schmidt-Gayk, H. (Heinrich), 1944. II. Armbruster,
F. P. (Franz Paul), 1957 . III. Bouillon, R.
[DNLM: 1. Biological Assay--methods. 2. Calcitonin--analysis.
3. Calcium--analysis. 4. Parathyroid Hormone--analysis.
5. Radioimmunoassay--methods. 6. Vitamin D--metabolism.
QV276 CI442] QP572.P3C35 1990 616.07'56--dc20 DNLMlDLC
for Library of Congress 90-10022
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Origina1ly published by Springer-Verlag Berlin Heidelberg New York in 1990.
Softcover reprint of the hardcover 1s t edition 1990
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List of Contributors
Aker, I., Clinical Laboratory, Department of Surgery, University of Heidelberg,
D-6900 Heidelberg, F.R.G.*
Armbruster, F. P., Immundiagnostik GmbH, Wilhelmstr. 7, D-6140 Bensheim
Bellet, D., Unite d'Immunochimie, Institut Gustave-Roussy, and Faculte des
Sciences Pharmaceutiques et Biologiques, Paris, France
Birringer, H., Clinical Laboratory, Department of Surgery, University of
Heidelberg, D-6900 Heidelberg, F.R. G. *
Blind, E., Department of Endocrinology, University of Heidelberg, D-6900
Heidelberg, F.R.G.
Bohler. U., Clinical Laboratory, Department of Surgery, University of Heidel
berg, D-6900 Heidelberg, F.R.G.*
Bohuon, C., Department of Clinical Biology, Institut Gustave-Roussy and
Faculte des Sciences Pharmaceutiques et Biologiques, Chatenay Malabry, France
Bothe, V., Clinical Laboratory, Department of Surgery, University of Heidel
berg, D-6900 Heidelberg, F.R.G.*
Bouillon, R., Katholieke Universiteit Leuven, Laboratorium voor Experimen
tele Geneeskunde en Endocrinologie, Onderwijs en Navorsing, Gasthuisberg,
B-3000 Leuven, Belgium
Fischer, S., Department of Surgery, Abt. 2.1.1, University of Heidelberg, D-6900
Heidelberg, F.R.G.
Flentje, D., Department of Surgery, Abt. 2.1.1, University of Heidelberg,
D-6900 Heidelberg, F.R.G.
Ghillani, P., Unitee d'Immunochimie, Institut Gustave-Roussy, F-94805
Villejuif Cedex, France
Grauer, A., Department of Endocrinology, University of Heidelberg, D-6900
Heidelberg, F.R.G.
Herfarth, K., Department of Surgery, Abt. 2.1.1, University of Heidelberg,
D-6900 Heidelberg, F.R.G.
Hitzler, W., Clinical Laboratory, Department of Surgery, University of Heidel
berg, D-6900 Heidelberg, F.R.G.
VI List of Contributors
Horst, R. L., National Animal Disease Center, U.S. Department of Agriculture,
Agricultural Research Service, P.O. Box 70, Ames, IA S0010, USA
Kunst, G., Clinical Laboratory, Department of Surgery, University of Heidel
berg, D-6900 Heidelberg, F.RG.*
Limbach, H. J., 1m Breitspiel15, D-6900 Heidelberg, F.R.G.
Mayer, E., LindenstraBe 17, D-S4S0 Neuwied 22, F.RG.
Motte, Ph., lnstitut Merieux, 1541 avenue Marcel Merieux, F-69280 Marcy
l'Etoile, France
Ratcliffe, J. G., Wolfson Research Laboratories, Department of Clinical
Chemistry, Queen Elizabeth Medical Centre, Birmingham B15 2TH, United
Kingdom
Ratcliffe, W. A., Wolfson Research Laboratories, Department of Clinical
Chemistry, Queen Elizabeth Medical Centre, Birmingham B1S 2TH, United
Kingdom
Raue, F., Department of Endocrinology, University of Heidelberg, D-6900
Heidelberg, F.R.G.
Reichel, H., Department of Nephrology, University of Heidelberg, D-6900
Heidelberg, F.RG.
Reinhardt, T. A., National Animal Disease Center, U.S. Department of Agri
culture, Agricultural Research Service, P.O. Box 70, Ames, IA S0010, USA
Scharfenstein, H., Clinical Laboratory, Department of Surgery, University of
Heidelberg, D-6900 Heidelberg, F.R.G.*
Scharla, S., Department of Endocrinology, University of Heidelberg, D-6900
Heidelberg, F.R.G.
Schmidt-Gayk, H., 1m Breitspiel15, D-6900 Heidelberg, F.R.G.
Schneider. H.-G., Department of Endocrinology, University of Heidelberg,
D-6900 Heidelberg, F.RG.
Stadler, A., Clinical Laboratory, Department of Surgery, University of Heidel
berg, D-6900 Heidelberg, F.RG.*
Troalen, F., Unite d'lmmunochimie, lnstitut Gustave-Roussy, F-9480S Villejuif
Cedex, France
Vogel, G., Clinical Laboratory, Department of Surgery, University of Heidel
berg, D-6900 Heidelberg, F.RG.*
Walch, S., 1m Breitspiel15, D-6900 Heidelberg, F.RG.
Zillikens, D., Clinical Laboratory, Department of Surgery, University of
Heidelberg, D-6900 Heidelberg, F.RG.*
* address all correspondense to the first editor.
Preface
There are very few books available containing details on the assay of calcium
regulating hormones. The volume Assay of Calcium Regulating Hormones,
edited by D. D. Bickle, was published in 1983, but since then rapid progress
has been made in our understanding of calcium matabolism and in the assays of
calcium regulating hormones. To mention just a few:
A revised structure of parathyroid hormone (PTH) has been confirmed by
cDNA techniques. The peptides with the correct structure are commercially
available, and assays for intact PTH have been developed that exhibit extreme
sensitivity and specificity.
Parathyroid hormone-related protein (PTH-rP) is a substance that has been
recently discovered. Assays for PTH-rP have been developed, but many que
stions remain to be answered about the clinical usefulness of the assayofPTH-rP.
The clinical significance of vitamin D metabolites in serum has become much
clearer, and more direct measurements of 25-hydroxyvitamin D and 1,25-dihy
droxyvitamin D allow an easier diagnosis and more appropriate therapeutic
intervention.
The diagnostic value of new peptides from the calcitonin gene has been investi
gated. Highly sensitive immunoradiometric assays for the measurement of calci
tonin in serum or plasma have recently been introduced, facilitating the diagnosis
of early familiar medullary thydroid carcinoma. It is hoped that the physiology
and pathophysiology of calcitonin may be clarified by these new assays.
We have felt the lack of a recent review on calcium regulating hormones and
related topics and have consequently attempted to provide a collection of clinical
reviews and detailed assay protocols. The first chapters deal with clinical in
formation relevant to calcium regulating hormones. These chapters are followed
by a description of methods for the assay of calcium regulating hormones, many
of which have already been described in other publications. As they are all up
dated, these second-generation assays replace older methods. Because some
assays have been in clinical use for years, it is now possible to evaluate their
clinical utility. We hope that the book will be welcomed by physicians and
chemists working in laboratories and clinics involved with calcium-related dis
eases and that it will be useful for years to come.
VIII Preface
We are pleased to have been able to cooperate with experts in endocrinology,
who have provided up-to-date constributions from their particular areas of ex
pertise. We also thank Springer-Verlag for its valuable assistance in preparing
the book.
Heidelberg, July 1990 H. Schmidt-Gayk
Contents
1 Introduction
1.1 Clinical Application of Intact Parathyroid Hormone Determination
(E. Blind, D. Flentje, and S. Fischer) . . . . . . . . . . . . . . . . . . . 3
1.2 Clinical Use of Vitamin D Metabolite Assays (Calcidiol and Calcitriol)
(R. Bouillon) ................................. 24
1.3 Diagnostic Value of the Peptides from the Calcitonin Gene (F. Raue,
H.-G. Schneider, and A. Grauer) . . . . . . . . . . . . . . . . . . . .. 48
1.4 Immunoassay Vial for Improved Assay Performance (H. Schmidt-
Gayk, H. J. Limbach, and S. Walch) ................... 60
2 Parathyroid Hormone Assays and Clinical Application
2.1 Homologous Radioimmunoassay for Human Parathyroid Hormone
(Residues 53--84) with Tyrosylated Peptide as Tracer
(W. Hitzler) .................................. 69
2.2 Sensitive Homologous Radioimmunoassay for Human Parathyroid
Hormone (Residues 53-84) to Diagnose Hypoparathyroid
Conditions: Guidelines for the Establishment of Highly Sensitive
Assays (D. Zillikens) . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 84
2.3 Rapid Homologous Radioimmunoassay for Human Parathyroid
Hormone (Residues 53-84) as a Diagnostic Aid in Hypercalcemic
Crisis (D. Zillikens) ............................. 96
2.4 Homologous Radioimmunoassay for Human Parathyroid Hormone
(Residues 44-68) with Tyrosylated Peptide as Tracer (H. Schmidt-
Gayk and G. Kunst) ............................. 107
2.5 Purification of 125Iodine-Labeled Tyrosylated Human Para-
thyroid Hormone Fragment (Residues 44-68), Optimized by Reverse
Phase High-Performance Liquid Chromatography (G. Vogel) .... 116
x Contents
2.6 Homologous Radioimmunoassay for Human Parathyroid Hormone
(Residues 28-48) with Tyrosylated Peptide as Tracer (I. Aker) . .. 127
2.7 Homologous Radioimmunoassay for Human Parathyroid Hormone
(Residues 1-34) with Biotinylated Peptide as Tracer (A. Stadler) 137
2.8 Measurement of Intact Parathyroid Hormone by an Extracting
Two-Site Immunoradiometric Assay (E. Blind) ............ 151
2.9 Measurement of Intact Parathyroid Hormone by a Two-Site
Immunochemiluminometric Assay (U. Bohler) ............ 167
2.10 A Sensitive Adenylate Cyclase Bioassay for Parathyroid Hormone
(H. Scharfenstein) . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 190
2.11 Immunological Properties of Asparagine versus Aspartic Acid at
Residue 76 of Human Parathyroid Hormone (Residues 53-84)
(K. Herfarth, E. Blind, H. Schmidt-Gayk, and F. P. Armbruster) . 203
3 Parathyroid Hormone-Related Protein
3.1 Parathyroid Hormone-Related Protein (W. A. Ratcliffe and
J. G. Ratcliffe) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 213
4 Cyclic AMP
4.1 Plasma and Urinary Cyclic AMP
Assay and Clinical Application (F. P. Armbruster) . . . . . . . . .. 231
5 Vitamin D: Assays of 25-Hydroxyvitamin D (Calcidiol) and
~25-Dihydroxyvitamin D (Calcitriol)
5.1 Simultaneous Determination of 25-Hydroxyvitamin D2 and 25-
Hydroxyvitamin D3 by High-Performance Liquid Chromatography
(E. Mayer and H. Schmidt-Gayk) .................... 247
5.2 Competitive Protein-Binding Assay for the Diagnosis of Hyper-
and Hypovitaminosis D (Y. Bothe and H. Schmidt-Gayk) . . . . .. 258
5.3 A Competetive Protein-Binding Assay with Second Antibody
Separation for the Diagnosis of Hyper- and Hypovitaminosis D
(H. Birringer and H. Schmidt-Gayk) .................. 280
5.4 Interlaboratory Comparison of Calcidiol Determination
(E. Mayer) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 286
Contents XI
5.5 A Sensitive Radioimmunoassay for 1,25-Dihydroxyvitamin D3
(Calcitriol) after High Performance Liquid Chromatography of
Plasma or Serum Extracts (S. Scharla and H. Reichel) . . . . . . .. 300
5.6 A Double Antibody Radioimmunoassay for measurement of
la, 25-Dihydroxyvitamin D3 (F. P. Armbruster, H. Reichel,
and H. Schmidt-Gayk) . . . . . . . . . . . . . . . . . . . . . . . . . .. 318
5.7 Competitive Protein-Binding Assay of Calcitriol with an Advanced
Preparation of Bovine Calf Thymus Cytosol (T. A. Reinhardt
and R. L. Horst) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 334
6 Assay of Calcitonin and Related Peptides
6.1 Design of New Methods for Measuring Calcitonin and Related
Peptides Using Monoclonal Antipeptide Antibodies (Ph. Motte,
P. Ghillani, F. Troalen, C. Bohoun, and D. Bellet) . . . . . . . . .. 349
Subject Index . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 365
