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Purification Of Purification And Purification

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When examining the purification data in Table 4, we can analyze the different methods of purification of LDH and determine how effective each methods is in order to ultimately purify LDH. Unfortunately, there are two different definitions of purification: one is have a high end yield of the protein, while the other is having a high end concentration without any contaminating proteins. Furthermore, in order to achieve one type of purification, the other one has to be given up. The very first purification step involved the 65% ammonium sulfate cut that resulted in a highest protein concentration out of all the purification steps. The protein concentration went up from 3.925mg/ml from the clarified homogenate to 28.11mg/ml, which is resulted of the pelleting out of the LDH and reducing the volume from 114mL to 5.7mL. Furthermore, the 65% pellet cut was able to recovery 46% of the LDH, while purifying it by 6.8 fold. Also, this step was able to remove 65% of the total proteins, while retaining 35% of it. Due to the increased purity and 46% recovery of LDH we can conclude that most of the removed total protein was non-LDH, contaminating protiens. Unfortunately, since this was only the first purification step, the amount of comtaminating proteins is still fairly high as depicted in Figure 14, where the 65% cut pellet contains several dark bands besides the one at 35kDa. The second purification step involved the affinity chromatography column that resulted in a sharp decrease
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