[医学]生物大分子分离纯化.pptVIP

* See slide 22. This is the data from the UNOsphere Q tech note. * * * * * Hydroxyapatite is an extremely versatile chromatography tool. It has been used for over thirty years for separation of biological molecules of all kinds ... it is used to separate and purify ... Hydroxyapatite is a crystalline calciumphosphate compound which is hydroxylated. Bio-Rad is the largest manufacturer of hydroxyapatite for chromatography in the world, we make several ton every years which are used in both laboratories and pharmaceutical and diagnostic production all over the world. * It is easy to see the difference between the old crystalline material and the new ceramic spherical material. * The new ceramic hydroxyapatite has all the unique separation properties of the crystalline material, but none of it’s limitations. * * * * * Fraction 27 (lane 6 on BRX SDSgel) from the BRX S column was injected onto a Bio-Scale CHT2-I column. The antibody was eluted with a combination NaCl/NaPO3 gradient (similar to the IgGT CHT application note) conditions. Notice the narrow peak! Not a bad recommendation for your CHT customers. Purity of the eluted antibody is shown on lanes 4 and 7 on CHT SDSgel (slide # 14). Profinity IMAC Resins 固定化金属螯合层析：Immobilized Metal Affinity Chromatography （IMAC） 基于纯化重组组氨酸标记蛋白设计，专门纯化His-tagged Protein 螯合带电离子，如Zn2+,Ni2+,Cu2+, 现有的产品为螯合Ni2+的介质 UNOsphere 技术 更好的流动特性，在高流速下载量，目的蛋白得率和纯度不会受到影响。 IDA 各种流速下的载量和分辨率 高流速，高载量，高分辨率，低反压 Profinity IMAC Resins 高流速、低反压 一般的Protocol Profinity IMAC Resins 上样缓冲液：20-50mM磷酸钠或磷酸钾缓冲液，含0.3－1M KCl，pH8.0 清洗缓冲液：上样缓冲液含有5－20mM咪唑，或上样缓冲液pH降到6.3后 进行清洗 洗脱缓冲液：20－500mM咪唑，如50mM磷酸钠缓冲液，含0.3M KCl, 0.5M 咪唑， pH8.0 或走5－500mM咪唑的线性梯度，20以上柱床体积，以确定最佳清洗和洗脱的咪唑浓度 不同 IMAC 介质纯化氨基肽酶(aminopeptidase)纯度比较 Profinity IMAC, IDA ligand Profinity IMAC Resins 介质再生Protocol和使用寿命测试 Strip 50mM EDTA, 50mM sodium phosphate, 300mM NaCl(pH7.5) Clean 50mM sodium acetate, 300mM NaCl(pH4.0) Charge