公开(公告)号：US07317187B2

公开(公告)日：2008-01-08

申请号：US11135490

申请日：2005-05-24

申请人： William R. Davidson ,  Bori Shushan ,  Peter Kovarik ,  Tom R. Covey ,  John D. Brennan

发明人： William R. Davidson ,  Bori Shushan ,  Peter Kovarik ,  Tom R. Covey ,  John D. Brennan

IPC分类号： B01D59/44

CPC分类号： H01J49/164 ,  H01J49/0413

摘要： Sol-gel derived monolithic silica columns containing entrapped dihydrofolate reductase were used for frontal affinity chromatography of small molecule mixtures. The output from the column combined with a second stream containing the matrix molecule (HCCA) and was directly deposited onto a conventional MALDI plate that moved relative to the column via a computer controlled x-y stage, creating a semi-permanent record of the FAC run. The use of MALDI MS allowed for a decoupling of the FAC and MS methods allowing significantly higher ionic strength buffers to be used for FAC studies, which allowed for better retention of protein activity over multiple runs.

摘要翻译： 使用含有截留二氢叶酸还原酶的溶胶 - 凝胶衍生的整体硅胶柱用于小分子混合物的正面亲和层析。 来自柱的输出与含有基质分子（HCCA）的第二流组合，并直接沉积到通过计算机控制的x-y阶段相对于柱移动的常规MALDI板上，形成FAC运行的半永久记录。 使用MALDI MS允许FAC和MS方法的去耦，从而允许显着更高的离子强度缓冲液用于FAC研究，这允许在多次运行中更好地保​​留蛋白质活性。