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Development of an immunoassay for tartrate-resistant acid phosphatase and its use in the monitoring of bone metabolism.

Chi Keung Cheung. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 219-251). / Chapter CHAPTER I --- LITERATURE REVIEW / Chapter 1 --- The structure of bone --- p.2 / Chapter 1.1. --- The cortical bone --- p.3 / Chapter 1.2. --- The cancellous bone --- p.3 / Chapter 2 --- The composition of bone --- p.3 / Chapter 2.1. --- Bone minerals --- p.4 / Chapter 2.2. --- The organic matrix --- p.4 / Chapter 2.3. --- The bone cells --- p.9 / Chapter 2.3.1. --- The osteoblast and the osteocyte --- p.9 / Chapter 2.3.2. --- The osteoclast --- p.11 / Chapter 3 --- Bone turnover - modelling and remodelling of bone --- p.13 / Chapter 3.1. --- Postulated sequence of bone remodelling --- p.14 / Chapter 4 --- Regulation of bone resorption --- p.16 / Chapter 4.1. --- Role of osteoblast and the lining cell on bone resorption --- p.17 / Chapter 5 --- Regulation of bone formation --- p.19 / Chapter 6 --- Effects of systemic hormones and local factors on bone metabolism --- p.20 / Chapter 6.1. --- Parathyroid hormone --- p.20 / Chapter 6.2. --- "1,25-dihydroxyvitamin D3" --- p.22 / Chapter 6.3. --- Calcitonin --- p.23 / Chapter 6.4. --- Prostaglandins --- p.23 / Chapter 6.5. --- Sex hormones --- p.24 / Chapter 6.6. --- Glucocorticoid --- p.26 / Chapter 6.7. --- Growth hormone --- p.27 / Chapter 6.8. --- Insulin --- p.28 / Chapter 6.9. --- Thyroid hormones --- p.29 / Chapter 6.10. --- Other systemic and local factors --- p.30 / Chapter 7 --- Indices of bone turnover --- p.34 / Chapter 8 --- Non-biochemical indices of bone metabolism --- p.34 / Chapter 8.1. --- Radionuclide bone scan --- p.34 / Chapter 8.2. --- Radiokinetic assessment --- p.35 / Chapter 8.3. --- Bone biopsy --- p.35 / Chapter 8.4. --- Bone densitometry --- p.36 / Chapter 9 --- Biochemical indices of bone metabolism --- p.37 / Chapter 10 --- Biochemical markers of bone formation --- p.38 / Chapter 10.1. --- Alkaline phosphatase --- p.38 / Chapter 10.1.1. --- Role and origin of bone alkaline phosphatase isoenzyme --- p.39 / Chapter 10.1.2. --- Measurement of bone alkaline phosphatase --- p.41 / Chapter 10.1.2.1. --- Heat inactivation --- p.42 / Chapter 10.1.2.2. --- Chemical inactivation --- p.43 / Chapter 10.1.2.3. --- Immunological methods --- p.44 / Chapter 10.1.2.4. --- High performance liquid chromatography --- p.45 / Chapter 10.1.2.5. --- Gel electrophoresis --- p.45 / Chapter 10.1.2.6. --- Isoelectric focusing --- p.47 / Chapter 10.2. --- Osteocalcin --- p.48 / Chapter 10.3. --- Osteonectin --- p.51 / Chapter 10.4. --- Matrix Gla-protein --- p.51 / Chapter 10.5. --- Other non-collagenous proteins --- p.52 / Chapter 10.6. --- Urinary Gla concentration --- p.52 / Chapter 10.7. --- Collagen peptides and extension peptides --- p.54 / Chapter 11 --- Biochemical markers of bone resorption --- p.55 / Chapter 11.1. --- Urine hydroxyproline --- p.55 / Chapter 11.2. --- Pyridinium cross-links --- p.58 / Chapter 11.3. --- Acid phosphatase --- p.60 / Chapter 11.3.1. --- Acid phosphatase isoenzymes --- p.60 / Chapter 11.3.2. --- The band 5 acid phosphatase isoenzyme genetics and characteristics --- p.62 / Chapter 11.3.3. --- Band 5 acid phosphatase as marker of osteoclastic function --- p.64 / Chapter 11.3.4. --- Measurement of osteoclastic acid phosphatase --- p.67 / Chapter 11.3.4.1. --- Specific chemical inhibitor --- p.67 / Chapter 11.3.4.2. --- Electrophoresis --- p.67 / Chapter 11.3.4.3. --- Immunological methods --- p.68 / Chapter 12 --- Problems with current biochemical markers of bone metabolism --- p.68 / Chapter 13 --- Aims of this study --- p.70 / Chapter CHAPTER II --- PURIFICATION OF TARTRATE-RESISTANT ACID PHOSPHATASE AND THE DEVELOPMENT OF AN IMMUNOASSAY FOR IT'S MEASUREMENT / Chapter 1 --- Introduction --- p.72 / Chapter 2 --- Materials and methods --- p.75 / Chapter 2.1. --- Chemicals and reagents --- p.75 / Chapter 2.1.1. --- Apparatus --- p.76 / Chapter 2.2. --- Methods --- p.77 / Chapter 2.2.1. --- Cord serum --- p.77 / Chapter 2.2.2. --- Measurement of tartrate-resistant acid phosphatase activity --- p.77 / Chapter 2.2.3. --- Measurement of protein concentration --- p.80 / Chapter 2.2.4. --- Purification of TRACP from cord plasma --- p.82 / Chapter 2.2.4.1. --- Cation-exchange column chromatography --- p.83 / Chapter 2.2.4.2. --- Gel filtration column chromatography --- p.84 / Chapter 2.2.4.3. --- Concanavalin A-affinity column chromatography --- p.85 / Chapter 2.2.4.4. --- Preparative isoelectric focusing (IEF) --- p.86 / Chapter 2.3. --- Characterisation of purified TRACP --- p.90 / Chapter 2.3.1. --- Polyacrylamide gel electrophoresis (PAGE) --- p.91 / Chapter 2.3.2. --- "Optimum pH, substrate specificity and the effects of potential activators and inhibitors on TRACP activity" --- p.99 / Chapter 2.3.3. --- Amino acid composition of purified TRACP --- p.101 / Chapter 2.4. --- Methods for raising anti-human TRACP antibody and characterisation of the antiserum --- p.102 / Chapter 2.4.1. --- Production of rabbit anti-human TRACP antibody --- p.102 / Chapter 2.4.2. --- Determination of the titre of rabbit anti-human TRACP antibody --- p.103 / Chapter 2.4.3. --- Immunoblotting analyses for cross reactivity study --- p.103 / Chapter 2.4.4. --- Immunohistochemical study for antibody specificity --- p.105 / Chapter 2.4.5. --- Cross reactivity study of the rabbit anti-human TRACP antibody to some tissue preparations --- p.107 / Chapter 2.5. --- Enzyme linked immunosorbent assay for TRACP --- p.109 / Chapter 2.5.1. --- Optimisation and evaluation of the new ELISA method for TRACP --- p.111 / Chapter 3 --- RESULTS --- p.113 / Chapter 3.1. --- "Precision of methods for the determination of protein, TRACP and phosphate." --- p.113 / Chapter 3.2. --- Isolation and purification of TRACP --- p.113 / Chapter 3.2.1. --- Concanavalin A affinity chromatography --- p.120 / Chapter 3.2.2. --- Isoelectric focusing (IEF) --- p.120 / Chapter 3.3. --- Characterisation and homogeneity of purified TRACP --- p.128 / Chapter 3.3.1. --- Characterisation of purified TRACP --- p.128 / Chapter 3.3.2. --- Homogeneity of purified TRACP --- p.132 / Chapter 3.3.3. --- Amino acid composition --- p.136 / Chapter 3.4. --- Characterisation of the rabbit anti-human TRACP antibody --- p.136 / Chapter 3.4.1. --- Antibody specificity - immunoblotting study --- p.139 / Chapter 3.4.2. --- Antibody specificity - cross reactivity with partially purified non-cord plasma TRACP --- p.142 / Chapter 3.4.3. --- Antibody specificity - immunohistochemical study --- p.145 / Chapter 3.5. --- Enzyme linked immunosorbent assay for TRACP --- p.145 / Chapter 3.5.1. --- Optimal concentration of antigen for coating of microtitre plate --- p.145 / Chapter 3.5.2. --- Kinetics of reaction with the primary rabbit anti-human TRACP antibody --- p.149 / Chapter 3.5.3. --- "Precision, recovery and assay range" --- p.149 / Chapter 4 --- DISCUSSION --- p.155 / Chapter 4.1. --- Purification of cord plasma TRACP --- p.155 / Chapter 4.2. --- Characterisation of cord plasma TRACP --- p.158 / Chapter 4.3. --- Characterisation of rabbit anti-human TRACP antibody --- p.163 / Chapter 4.4. --- Enzyme immunoassay for TRACP --- p.165 / Chapter CHAPTER III --- STUDY OF SERUM TRACP IN HEALTHY SUBJECTS AND IN PATIENTS WITH BONE RELATED DISEASES / Chapter 1 --- Introduction --- p.168 / Chapter 2 --- Materials and methods --- p.171 / Chapter 2.1. --- Subjects --- p.171 / Chapter 2.1.1. --- Healthy subjects --- p.171 / Chapter 2.1.2. --- Patients --- p.172 / Chapter 2.1.2.1. --- Post-menopausal women on hormone replacement therapy --- p.172 / Chapter 2.1.2.2. --- Hip fracture patients --- p.173 / Chapter 2.1.2.3. --- Other patients --- p.174 / Chapter 2.3. --- Measurement of other biochemical parameters --- p.175 / Chapter 2.3.1. --- Bone alkaline phosphatase --- p.175 / Chapter 2.3.2. --- "Measurement of urine hydroxyproline, creatinine, calcium, osteocalcin, thyroid hormones and parathyroid hormone" --- p.176 / Chapter 2.4. --- Statistics --- p.178 / Chapter 3 --- RESULTS --- p.179 / Chapter 3.1. --- Healthy subjects --- p.179 / Chapter 3.2. --- Serum TRACP concentration in post-menopausal women before and after hormone replacement therapy --- p.185 / Chapter 3.3. --- TRACP concentration in elderly subjects with hip fractures --- p.189 / Chapter 3.4. --- Serum TRACP concentrations in patients with other bone related diseases --- p.190 / Chapter 3.4.1. --- Hyperthyroidism --- p.194 / Chapter 3.4.2. --- Hyperparathyroidism --- p.198 / Chapter 3.4.3. --- Haemodialysis --- p.201 / Chapter 4 --- DISCUSSION --- p.204 / GENERAL DISCUSSION --- p.216 / REFERENCES --- p.219

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_319240
Date January 1993
ContributorsCheung, Chi Keung., Chinese University of Hong Kong Graduate School. Division of Clinical and Pathological Sciences.
PublisherChinese University of Hong Kong
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xv, 251 leaves : ill. (chiefly mounted col.) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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