Taylor G A, Mazhindu H N, Findlay J B, Peacock M
Clin Chim Acta. 1986 Feb 28;155(1):31-41. doi: 10.1016/0009-8981(86)90096-3.
Vitamin D binding protein/group-specific component was purified from human plasma by chromatographic techniques utilising high performance liquid chromatography and by traditional low pressure chromatographic techniques alone. Use of high performance liquid chromatography considerably reduced the time taken to prepare pure vitamin D binding protein and increased the yield to 16% compared with 2.8% using the traditional methods. The vitamin D binding protein prepared by high performance liquid chromatography was shown to be highly pure by amino acid sequence, SDS gel electrophoresis and by antibody production. The amino acid sequence was confirmed and extended. The affinity constants of the high pressure liquid chromatography purified vitamin D binding protein for 25 hydroxycholecalciferol (25 OHD3) and 1,25 dihydroxycholecalciferol 1,25(OH)2D3 were 1.9 X 10(7) mol/l and 2.6 X 10(6) mol/l, respectively.
利用高效液相色谱和传统低压色谱技术从人血浆中纯化维生素D结合蛋白/组特异性成分。与传统方法相比，高效液相色谱的使用大大减少了制备纯维生素D结合蛋白所需的时间，并将产率提高到16%，而传统方法的产率为2.8%。通过氨基酸序列、SDS凝胶电泳和抗体产生证明，用高效液相色谱制备的维生素D结合蛋白高度纯净。氨基酸序列得到确认并有所扩展。高压液相色谱纯化的维生素D结合蛋白对25-羟基胆钙化醇（25-OHD3）和1,25-二羟基胆钙化醇1,25(OH)2D3的亲和常数分别为1.9×10(7) mol/l和2.6×10(6) mol/l。
