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

by Chak Chi Wai. / Thesis (M.Phil)--Chinese University of Hong Kong, 1992. / Includes bibliographical references (leaves 142-155). / ACKNOWLEDGMENTS --- p.i / TABLE OF CONTENT --- p.ii / LIST OF ABBREVIATION --- p.viii / ABSTRACT --- p.x / Chapter CHAPTER ONE: --- INTRODUCTION / Chapter 1.1 --- INTRODUCTION TO ALKALINE PHOSPHATASE --- p.2 / Chapter 1.1.1 --- The Alkaline Phosphatase Isoenzymes --- p.2 / Chapter 1.1.2 --- The Properties of Alkaline Phosphatases --- p.5 / Chapter 1.1.3 --- Serum Alkaline Phosphatases --- p.7 / Chapter 1.1.3.1 --- Intestinal Alkaline Phosphatase --- p.8 / Chapter 1.1.3.2 --- Placental Alkaline Phosphatase --- p.8 / Chapter 1.1.3.3 --- Renal Alkaline Phosphatase --- p.9 / Chapter 1.1.3.4 --- Skeletal Alkaline Phosphatase --- p.9 / Chapter 1.1.3.5 --- Hepatic Alkaline Phosphatase --- p.10 / Chapter 1.1.3.6 --- Miscellaneous Alkaline Phosphatases --- p.10 / Chapter 1.1.4 --- Problems in Discriminating the Skeletal and Hepatic Alkaline Phosphatase in Serum --- p.12 / Chapter 1.1.5 --- Wheat Germ Lectin Precipitation of the Bone- Specific Alkaline Phosphatase --- p.13 / Chapter 1.2 --- STRUCTURE OF BONE AND MECHANISMS OF CALCIFICATION --- p.15 / Chapter 1.2.1 --- Gross Structure of Bone --- p.15 / Chapter 1.2.2 --- The Elements of Bone --- p.17 / Chapter 1.2.2.1 --- Bone Cells --- p.17 / Chapter 1.2.2.2 --- Organic Substances of Bone --- p.19 / Chapter 1.2.2.3 --- Inorganic Substances of Bone --- p.21 / Chapter 1.2.3 --- Mechanisms of Calcification --- p.22 / Chapter 1.3 --- BONE FRACTURE HEALING --- p.24 / Chapter 1.3.1 --- Types of Fracture --- p.24 / Chapter 1.3.2 --- The Process of Bone Fracture Healing --- p.26 / Chapter 1.3.2.1 --- Stage of Hematoma --- p.26 / Chapter 1.3.2.2 --- Stage of Subperiosteal and Endosteal Cellular Proliferation --- p.28 / Chapter 1.3.2.3 --- Stage of Fibrocartilaginous Callus --- p.28 / Chapter 1.3.2.4 --- Stage of Bony Callus --- p.30 / Chapter 1.3.2.5 --- Stage of Remodeling --- p.31 / Chapter 1.4 --- THE OSTEOBLASTIC CHARACTERS OF UMR-106 OSTEOSARCOMA CELL LINE --- p.32 / Chapter 1.4.1 --- Classification of Osteosarcoma --- p.32 / Chapter 1.4.2 --- Derivation of UMR-106 Osteosarcoma Cell Line --- p.33 / Chapter 1.4.3 --- Osteoblastic Characters of UMR-106 --- p.34 / Chapter 1.4.3.1 --- ALP Expression --- p.34 / Chapter 1.4.3.2 --- Hormone Responsive Adenylate Cyclase System --- p.35 / Chapter 1.4.3.3 --- "Cytosolic Receptors for 1,25-Dihydroxy- cholecalciferol" --- p.35 / Chapter 1.5 --- IN VITRO CULTURE OF FETAL RAT CALVARIAL OSTEOBLASTS --- p.37 / Chapter 1.6 --- AIM AND SCOPE OF THIS DISSERTATION --- p.39 / Chapter CHAPTER TWO: --- MATERIALS AND METHODS / Chapter 2.1 --- BONE FRACTURE OPERATION --- p.42 / Chapter 2.1.1 --- Animals --- p.42 / Chapter 2.1.2 --- Blood Sampling and Preparation of Plasma Samples --- p.42 / Chapter 2.1.3 --- Bone Fracture Operation --- p.43 / Chapter 2.1.3.1 --- Reagents and Apparatus --- p.43 / Chapter 2.1.3.2 --- Procedures --- p.44 / Chapter 2.1.4 --- Radiography --- p.50 / Chapter 2.1.5 --- Removal of Tibiae --- p.51 / Chapter 2.1.6 --- Extraction of Callus ALP --- p.51 / Chapter 2.1.6.1 --- Reagent --- p.51 / Chapter 2.1.6.2 --- Homogenization of the Callus --- p.51 / Chapter 2.1.6.3 --- Extraction of ALP --- p.52 / Chapter 2.1.7 --- Assay for Bone-Specific ALP --- p.53 / Chapter 2.1.7.1 --- Reagents --- p.53 / Chapter 2.1.7.2 --- Procedures --- p.54 / Chapter 2.1.8 --- Normal Curve for Plasma Bone-Specific ALP in Rabbits --- p.56 / Chapter 2.1.9 --- The Effects of Tibial Fracture on the Plasma Level of Bone-Specific ALP in Rabbits --- p.56 / Chapter 2.1.10 --- Profile of Plasma Bone-Specific ALP upon a Fracture Healing --- p.57 / Chapter 2.1.11 --- Profile of Callus Bone-Specific ALP at Different Stages of Fracture Healing --- p.57 / Chapter 2.2 --- CLINICAL STUDIES OF PLASMA BONE-SPECIFIC ALP --- p.58 / Chapter 2.2.1 --- Patient Groups --- p.58 / Chapter 2.2.1.1 --- Normal Adults --- p.58 / Chapter 2.2.1.2 --- Fracture Group --- p.58 / Chapter 2.2.1.3 --- Tumor Group --- p.59 / Chapter 2.2.2 --- Assays for Plasma Bone-Specific ALP --- p.59 / Chapter 2.3 --- "IN VITRO CULTURES OF FETAL, RAT OSTEOBLASTS AND UMR-106 OSTEOSARCOMA cell line" --- p.60 / Chapter 2.3.1 --- Animals --- p.60 / Chapter 2.3.2 --- UMR-106 Cell Line --- p.60 / Chapter 2.3.3 --- General Reagents Used for Cell Culture --- p.60 / Chapter 2.3.4 --- Isolation of Calvarial Osteoblasts --- p.64 / Chapter 2.3.4.1 --- Tools and Reagents --- p.64 / Chapter 2.3.4.2 --- Procedures --- p.65 / Chapter 2.3.5 --- Storage of UMR-106 Cell Line --- p.67 / Chapter 2.3.6 --- Subculture of Confluent Monolayer --- p.68 / Chapter 2.3.6.1 --- Reagents --- p.68 / Chapter 2.3.6.2 --- Procedures --- p.69 / Chapter 2.3.7 --- Staining for Calcium Deposits --- p.69 / Chapter 2.3.7.1 --- Reagents --- p.70 / Chapter 2.3.7.2 --- Procedures --- p.70 / Chapter 2.3.8 --- Protein Determination --- p.71 / Chapter 2.3.8.1 --- Reagents --- p.71 / Chapter 2.3.8.2 --- Procedures --- p.71 / Chapter 2.3.9 --- Microdetermination of Inorganic Phosphate --- p.72 / Chapter 2.3.9.1 --- Reagents --- p.72 / Chapter 2.3.9.2 --- Procedures --- p.73 / Chapter 2.3.10 --- Determination of Calcium --- p.73 / Chapter 2.3.10.1 --- Reagent --- p.73 / Chapter 2.3.10.2 --- Procedures --- p.73 / Chapter 2.3.11 --- Extraction and Assay for Cellular ALP --- p.74 / Chapter 2.3.11.1 --- Reagents --- p.74 / Chapter 2.3.11.2 --- Procedures --- p.75 / Chapter 2.3.12 --- Cell Surface ALP Assay --- p.75 / Chapter 2.3.12.1 --- Reagents --- p.75 / Chapter 2.3.12.2 --- Procedures --- p.76 / Chapter 2.3.13 --- Extraction of Calcium Phosphate Deposits --- p.76 / Chapter 2.3.13.1 --- Reagent --- p.76 / Chapter 2.3.13.2 --- Procedures --- p.76 / Chapter 2.3.14 --- Collagen Synthesis Assay --- p.77 / Chapter 2.3.14.1 --- Reagents --- p.77 / Chapter 2.3.14.2 --- Procedures --- p.78 / Chapter CHAPTER THREE: --- EFFECTS OF TIBIAL FRACTURE ON THE LEVEL OF BONE-SPECIFIC ALKALINE PHOSPHATASE IN RABBITS / INTRODUCTION --- p.81 / results: / Chapter 3.1 --- normal curve for plasma bone-specific alp in rabbits --- p.82 / Chapter 3.2 --- THE EFFECTS OF TIBIAL FRACTURE ON THE PLASMA LEVEL OF BONE-SPECIFIC ALP IN RABBITS --- p.84 / Chapter 3.3 --- PROFILE OF THE PLASMA ALP LEVEL UPON HEALING OF TIBIAL FRACTURE --- p.86 / Chapter 3.4 --- RADIOGRAPHY --- p.89 / Chapter 3.5 --- PROFILE OF CALLUS BONE-SPECIFIC ALP ACTIVITY UPON HEALING OF TIBIAL FRACTURE --- p.93 / DISCUSSION --- p.95 / Chapter CHAPTER FOUR: --- CLINICAL STUDIES OF PLASMA BONE-SPECIFIC ALKALINE PHOSPHATASE / INTRODUCTION --- p.100 / RESULTS: / Chapter 4.1 --- NORMAL VALUES --- p.100 / Chapter 4.2 --- FRACTURE GROUP --- p.101 / Chapter 4.3 --- BONE TUMOR GROUP --- p.102 / DISCUSSION --- p.102 / Chapter CHAPTER FIVE: --- IN VITRO CULTURE OF FETAL RAT OSTEOBLASTS AND UMR-106 CELL LINE / INTRODUCTION --- p.105 / RESULTS: / Chapter 5.1 --- IN VITRO MINERALIZATION OF UMR-106 CELLS AND PRIMARY RC CELLS --- p.107 / Chapter 5.2 --- STUDY OF BONE-SPECIFIC ALP RELEASED INTO MEDIUM BY UMR-106 CELLS AND PRIMARY RC CELLS --- p.113 / Chapter 5.3 --- STUDY OF CELLULAR ALP ACTIVITIES AND CALCIUM PHOSPHATE DEPOSITS --- p.116 / Chapter 5.4 --- STUDIES OF CELLULAR ALP ACTIVITIES AND RELATIVE RATES OF COLLAGEN SYNTHESIS --- p.125 / DISCUSSION --- p.128 / Chapter CHAPTER SIX: --- GENERAL DISCUSSION --- p.136 / BIBLIOGRAPHY --- p.142 / APPENDIX --- p.156

骺軟骨板是位於長骨骨骺的一個軟骨結構，其主要功用為使骨骼延伸生長。 若人類骺軟骨板基因出現障礙，骨骼生長往往會受到嚴重的影響，使骨骼變得短小、畸形，造成殘障。此外，一些後天的系統性失調也會損害骺軟骨板的正常運作，導致矮小症。現今常用來醫治骨骼生長障礙的包括重組的人類生長荷爾蒙。然而，它的功效並非顯著，亦帶來很多的副作用；因此，我們希望能尋求更好的醫治方法。 / 近有的研究結果顯示，很多的內分泌及分泌因子能夠刺激骺軟骨板進行軟骨增生。但是，研究者卻往往未能將這些因子轉化及運用作醫療用途，原因是它們通常都是局部性地於骺軟骨板產出及發生效用。倘若以上提及的生長因子能給骺軟骨板的靶子蛋白鏈準確地被帶往骺軟骨板，那麼這些因子便能有效地被利用作治療用途，而其效能亦會給大大提高，副作用也會被減低。因此，我們現在進行研究的首要目標為於採用噬菌體展示和酵母展示方法， 尋找那些能認辨出骺軟骨板的靶子蛋白鏈及單鏈抗體。 / 於噬菌體展示庫裡，我們找出了兩條能認辨出小鼠骨骼軟骨細胞的靶子蛋白鏈 － C1 (RLDPTSYLRTFW) 和 C19 (HDSQLEALIKFM)。然而於體外測試實驗中， 它們對軟骨細胞及細胞外基質只擁有中度的親近性和特異性。此後，於酵母展示庫裡，我們亦發現三條可認辨某骺軟骨板蛋白的單鏈抗體 － 它們的親近性甚高 (達至1 nM)，而其對軟骨組織的特異性也為良好 (它們只認辨軟骨組織，但卻沒能認辨出其他的軟組織，包括腦、心臟、肝臟、肺臟、腎臟、脾臟、小腸及肌肉)。 其後，於小鼠胚胎免疫染色測試實驗中，這些單鏈抗體只亦選擇地認辨軟骨組織。再者，當這些單鏈抗體被注射入小鼠的靜脈中，它們也會準確地停留在軟骨組織裡，顯示出其特異性於體內測試實驗中亦存在。 / 總括而言，利用噬菌體展示和酵母展示方法，我們發現了一些能認辨出骺軟骨板的靶子蛋白鏈及單鏈抗體。這些單鏈抗體擁有對軟骨組織非常高的親近性和特異性。我們預計，若然將這些單鏈抗體和內分泌及分泌因子連結在一起，它們或能成為醫治骨骼生長障礙的新方法。 / The growth plate is a specialized cartilage structure at the ends of long bones that is responsible for bone elongation. Many human genetic disorders of the growth plate impair skeletal growth, often resulting in bones that are severely short and malformed, causing serious disability. Many acquired systemic disorders also impair growth plate function, causing short stature. Currently, recombinant human growth hormone is used to treat growth disorders, but it has limited efficacy for severe diseases and causes significant adverse effects. / Recent studies have identified many endocrine and paracrine factors that are capable of promoting chondrogenesis at the growth plate. However, the development of these molecules into effective therapies has been hampered by their action mechanism; they are produced locally and act locally in the growth plate and thus fail to lend themselves to systemic therapeutic approaches. We envisioned that, if these growth factors could be linked to growth plate-targeting peptides or polypeptides and then administered systemically, these molecules might provide a better treatment strategy for growth disorders because targeting might augment the therapeutic effect on the growth plate but diminish undesirable effects on non-target tissues. Toward this goal, we sought to identify peptides and antibody fragments that bind to cartilage tissue using phage display and yeast display technologies. / We used a phage display library that expressed linear peptides of 12 random amino acids on the phage surface and then selected for binding to murine primary cultured chondrocytes. This approach successfully identified two peptides, namely C1 (RLDPTSYLRTFW) and C19 (HDSQLEALIKFM), which exhibited moderate binding affinity and specificity to chondrocytes and extracellular matrix in vitro. We also used a yeast display library to identify three single-chain variable(scFv) antibody fragments that bound strongly to a growth plate-specific protein(EC50 values less than 1 nM). These antibody fragments demonstrated specific binding in vitro to homogenates of cartilage tissues, but not homogenates of brain, heart, liver, lung, kidney, spleen, small intestine, or muscle. Moreover, they also exhibited tissue-specific binding to cartilage structures in sections of mouse embryos. When these purified antibody fragments were injected intravenously in mice, they localized to cartilage and were not detectable in other tissues, including brain, heart, liver, lung, kidney, spleen, small intestine, or muscle, indicating that the antibody fragments were capable of specifically targeting cartilage tissue in vivo. / In conclusion, we employed phage display and yeast display methods to identify peptides and antibody fragments that bind to cartilage tissues. The selected antibody fragments showed particularly high binding affinity and specificity to cartilage. Conjugating these antibody fragments to endocrine and paracrine signaling molecules has the potential to provide targeted therapy for growth plate disorders. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Cheung, Sao Fong. / Thesis (Ph.D.) Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 89-102). / Abstracts also in Chinese.