Comparison of different types of ceramic hydroxyapatite for the chromatographic separation of plasmid DNA and a recombinant anti-Rhesus D antibody
Comparison of different types of ceramic hydroxyapatite for the chromatographic separation of plasmid DNA and a recombinant anti-Rhesus D antibody
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DOI：
10.1023/a:1011174905511
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发表时间：
2000-01-01
期刊：
影响因子：
--
通讯作者：
Freitag, R
中科院分区：
文献类型：
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作者：
Giovannini, R;Freitag, R
比较了三种陶瓷羟基磷灰石（HAP）的色谱性能。所有这三种材料都是通过烧结原始沉淀物制备的，尽管是在不同的温度下（I型、II型和III型HAP分别为400、700和1000 ℃）。这三种材料的孔径和压力限值不同（I型最低，III型最高）。I型和II型HAP的平均粒径为20 μ m。III型材料的粒度为40 μ m。将HAP-珠浆状填充到4 × 25-mm不锈钢柱中，并研究从澄清的E.大肠杆菌裂解物和来自CHO细胞培养物上清液的重组人抗体。三种HAP的色谱性能有明显差异，这与其对不同大小（4.7、10.3和11.4kb）的质粒和不同等电点蛋白质（溶菌酶，pI = 10.5;抗RhD抗体，pI = 8.3; β-乳球蛋白，pI = 4.9）的结合能力有关。吸附表面（孔径）的可及性，但也HAP表面上的结合位点的类型（P/C-网站比）提出的色谱行为的决定因素。
The chromatographic properties of three types of ceramic hydroxyapatite (HAP) are compared. All three materials were prepared by sintering the original precipitate, albeit at different temperatures (400, 700 and 1000 degreesC for type I, II and III HAP, respectively). The three materials differed in pore size and pressure limits (both lowest for type I and highest for type III). Type I and II HAP had an average particle diameter of 20 mum. The particle size of the type III material was 40 mum. HAP-beads were slurry-packed into 4 x 25-mm stainless steel columns and investigated for the chromatographic isolation of plasmid DNA from clarified E. coli lysates and of a recombinant human antibody from CHO cell culture supernatants respectively. The chromatographic performance of the three types of HAP showed significant differences, which were correlated to the binding capacities of the materials for (linearized) plasmids of different size (4.7, 10.3 and 11.4 kb) and proteins of different isoelectric point (lysozyme, pI = 10.5; anti RhD antibody, pI = 8.3; beta -lactoglobulin, pI = 4.9). The accessibility of the adsorptive surface (pore size) but also the types of binding sites on the HAP-surface (P/C-site ratio) are proposed as determining factors for the chromatographic behavior.