牛骨钙素的分离、纯化和放射免疫分析法的建立  

Separation and Purefied of Calf Osteocalcin，Set up Radioimmunoassay for Osteocalcin

作　　者：孔彦平[1] 孟迅吾[1] 周学瀛[1] 刘怀成[1] 刘书勤[1] 

机构地区：[1]中国医学科学院中国协和医科大学协和医院,北京100730

出　　处：《中国医学科学院学报》1997年第4期257-262,共6页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金!39370747;中国医学科学院科研基金!91006

摘　　要：采用凝胶过滤、快速液相（FPLC）和高压液相（HPLC）成功地从小牛股骨提取和纯化了骨钙素（BGP）。用其作为抗原免疫家免产生了高效价的抗体，38．5％结合率（Bo／T）的最终稀释度为1：35000。应用此抗体建立的放射免疫分析法，稳定性好，灵敏度高，亲和常数为2．29×1011，回收率为（102±2．2）％，批内变异为2．1％，批间变异为5．6％，同降钙素、甲状旁腺激素和胰岛素无任何交叉反应。81名健康志愿者血清BGP水平为（5．3±1．5）ng／ml。绝经后骨质疏松、原发性甲状旁腺功能亢进症、慢性肾功能不全和畸形性骨炎患者血清BGP值均明显高于健康者。而库欣（Cushing）综合征和甲状旁腺功能减退患者血清BGP水平明显低于健康者。本研究结果表明，骨转换增加的疾病，血BGP值有增高；骨形成降低或者转换减慢的疾病，其血BGP值有下降。因此，血清BGP浓度是反映骨转换敏感而可靠的指标，在骨代谢疾病的临床和科研中具有重要意义。Objective and Methods We extracted and purified osteocalcin（BGP） from calf femur bygel filtration， FPLC and HPLC subsequently． The rabbits were immunized by this antigen and produced antibody． With this antibody（final 1： 35 000 dilution） We established RIA for BGP． This assay is stable and sensitive， the kd was 2． 29×10 11．The intra assay variation was 2． 1％ and the inter assay variaton was 5． 6％． It has no crossreaction with Insulin，parathyroid hormone or calcitonin．Results The serum BGP values were detected in normal subjects and six kinds of bone metabolic diseases． In 81 normal subjects， the average serum BGP level was（5．3±1．5）ng／ml and in postmenopausal osteoporosis the serum BGP level [（6．2±1．9）ng／ml，n=42] was higher than the normal（P＜0．05）． The serum BGP levels were increased much in primary hyperparathyroidism [（12．8±7．0）ng／ml， n=21，P＜0．001]． Increased values of BGP were also found in patients with chronic renal failure [（8．5±2．4）ng／ml，n＝18，P＜0．01] and Paget' s disease [（6．7±2．2）ng／ml，n＝12，P＜0．05]．Decreased serum BGP values Were found in Cushing syndrome [（2．9±0．9）ng／ml， n =16， P＜0． 001」 and hypoparathyroidism [（3．4±1． 1） ng／ml， n=42，P＜0．001]． This re－sult showed that In diseases with high bone turnover， the serum BGP levels were Increased and in diseases with low bone turnover， the serum BGP levels were decreased． Conclusion The data suggest that serum BGP is a sensitive marker for bone turnover． It is very improtant in studying bone metabolic diseases．

关 键 词：骨钙素 提取 放射免疫法 骨质疏松 

分 类 号：Q513[生物学—生物化学] R681.02[医药卫生—骨科学]