Osteocalcin, also known as Bone-Gla-Protein (BGP), is a single chain (Mr=5800) vitamin K dependent protein produced by osteoblasts and found in high concentrations in bone (1-4). Post-translational modification by a vitamin K dependent carboxylase produces three g-carboxyglutamic acid residues at positions 17, 21 and 24. The mature protein contains 49 amino acids and a single intrachain disulfide bond joining Cys23 to Cys29. The secondary structure is highly calcium dependent and contains 14% α-helix, 20% β-sheet and 67% random form in the presence of calcium, and 1% α-helix, 20% β-sheet and 79% random form in the absence of calcium (4).
BGP binds to phospholipid vesicles in the presence of calcium ions (Kd=6 x 10-6 M) (5). BGP also binds hydroxyapatite and is an efficient inhibitor of hydroxyapatite seeded crystal growth, suggesting a regulatory role in bone mineralization (6). Although the fundamental role of BGP in vivo remains elusive, in vitro experimentation suggests roles in both bone resorption and bone formation. BGP has been shown to function as a chemo-attractant in an in vitro macrophage mediated bone resorption system. The serum concentration of BGP has been linked to bone formation rates, and BGP levels are currently used in research assessment of osteoporosis and other metabolic bone disease states (7).
Osteocalcin is prepared from bovine bone generally as described by Price et al. (1). This involves demineralization in EDTA, size exclusion and ion exchange chromatography. The purified protein is supplied 50 % (vol/vol) glycerol/0.01 M Tris, 0.075 M NaCl, pH 7.4, and should be stored at -20°C. Human osteocalcin is prepared from extracts of acid-demineralized bone, using immunoaffinity chromatography. The human protein is supplied in 0.02 M Tris, 0.15 M NaCl, 2 mM CaCl2, pH 7.4 and should be stored at -80°C. Purity is determined by SDS-PAGE.
Haemtech Biopharma可以运行多种分析技术，以提供对蛋白质药物产品或物质的生化分析，或帮助进行污染物鉴定。我们的测定，测试和技术菜单包括：应用领域IVIG药物的血栓形成性过程中杂质分析与止血有关的抗药物抗体测试（免疫原性）例如抗凝血因子抗体宿主细胞蛋白质的鉴定，定量和缓解在制品和在制品中药品的稳定性和释放测试与止血相关的重组蛋白的效力和纯度测定血浆蛋白分析蛋白质的结构表征翻译后修饰二硫键映射构象变化凝血测定试剂（例如凝血活酶）的表征定制的凝血产品检测应用药代动力学研究分析鉴定/验证工艺/产品验证蛋白质纯化方法的发展强迫降解研究方法高效液相色谱反相（RPC）大小排除（SEC）离子交换（IEC）疏水相互作用（HIC）金属亲和力（IMAC）蛋白质A（用于IVIG分析）SDS页面减少和不减少一维和二维考马斯亮蓝染色银染荧光染色免疫印迹多重分析定量化凝血测定因子分析（II，V，VII，VIII，IX，X，XI，XII，XIII等）PTPTPT行动计划福尔摩斯测试其他ELISA法各种显色测定因子分析（II，V，VII，VIII，IX，X，XI，XII，XIII等）其他翻译后修饰分析糖基化，硫酸化，磷酸化，羟基化，脱酰胺，乙酰化，氧化，氨基甲酸酯化和γ-羧化凝血酶生成测定对于IVIG和其他抗药物抗体ELISA（ADA）抗原料药抗体抗宿主细胞蛋白的抗体抗污染物或分解产物的抗体鉴定未知质谱N端测序残留水分测定顶空真空测量pH值渗透压折光率总蛋白质配方评估
