TAILIEUCHUNG - Biomimetic Based Applications Part 5

Tham khảo tài liệu 'biomimetic based applications part 5', kỹ thuật - công nghệ, cơ khí - chế tạo máy phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | The Improvement of LC-MS MS Proteomic Detection with Biomimetic Affinity Fractionation 151 4. Prefractionation of of rat liver cytosol proteins utilizing cascade composition of affinity chromatography Cascade affinity prefractionation of tissue samples From biomimetic affinity ligands library in our lab we first screened out some affinity ligands showing large absorbance differences in band distribution to rat liver cytosol proteins. Then the affinity ligands having medium absorbance ability were selected cascade composition of these affinity ligands could be adopted for the prefractionation of tissue proteins. In this study cascade composition of three affinity ligands was adopted and the schematic illustration of the workflow was shown in FIG. 6. Briefly after complex tissue sample was loaded to the first affinity ligand column flowthrough and elution were collected for the two new fractions. Then after above two new fractions were parallel loaded to the second affinity liand column flowthrough and elution were collected for the four new fractions. At last above four new fractions were loaded to the third affinity ligands and 8 new fractions could be obtained. The binding and elution conditions were same as above. After being desalted and concentrated using Microcon ultrafiltration membranes 3 000 Nominal Molecular Weight Limit Millipore all the fractions collected were used for subsequent SDS-PAGE electrophoresis and LTQ-MS MS analysis. Collecting all fractions LTQ-MS MS Fig. 6. Schematic illustration of Cascade affinity fraction used in this work 152 Biomimetic Based Applications Evaluating for differential protein absorbance profiles of mimetic affinity ligands After analyzing the screening data of our lab in serum and leech proteins we selected the affinity ligands having medium absorbance ability. Sixty affinity ligands were selected from the affinity ligands library of our lab and the protein absorbance characterization of these ligands was .

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