The Process of Protein Purification Literature review

The Process of Protein Purification - Literature review Example Vaccines, gene therapy and replacement (such as that in insulin-deficiency) all have helped in improving health conditions, and have been based on good elucidation of structures, research on structure-function relationships, and establishment of protein purification specific to the amino acid sequence present. With this in mind, this particular study designed a protocol to purify and characterize the a synthesis of cytochrome oxidase (SCO)-1-like protein 3966 in Streptomyces lividans. As will be seen later, better understanding of SCO proteins is still warranted, as many potential functions of these types of proteins are unclear. Moreover, SCO is a vital enzyme as the cytochrome oxidase c, and in essence the electron chain transport of the mitochondrial respiration mechanism, depends on it. Initial studies of homologues in bacteria have been the usual first step in protein characterization. Many proteins in the eukaryotic cells have been proven to have functional and structural counterparts in bacterial cells. Because of the relative ease of bacterial replication and protein purification, it is thus a method of choice in conducting in depth studies of proteins. REVIEW OF RELATED LITERATURE I. Protein Purification There are factors to consider in doing protein purification. ... There are many kinds of column chromatography, ion-exchange, affinity, and size exclusion are just some of the more usual protein purification procedures that may be done. Affinity chromatography uses antibodies for a specific protein as part of the column through which the protein solution passes. Although it is highly specific, it is more expensive and much harder to prepare. Size exclusion, on the other hand, depends on the differences of molecular weights of the proteins that are present in the solution. In general, proteins with high molecular weight are eluted fastest as they are not able to get into the small spaces of the column, making their path down the column less impeded. On the other hand, low molecular weight proteins still pass through the tiny spaces within the column, thus slowing down their descent. Although much easier to prepare than an affinity column, a size exclusion chromatography column is less specific, as different proteins of similar weigh are eluted out at the same time, despite them having differences in characteristics, such as the isoelectric point (Burgess, 2008).. For ion exchange chromatography, these beads are charged, thus attracting the oppositely-charged proteins present in the solution to be passed through the column. Depending on the objective of the experiment, the eluent or the bound proteins are collected for further processing such as concentrating. To get the proteins bound on the beads, salt solutions of graded concentrations are passed onto the column. As the concentration of the salt increases, the beads will more likely bind to the salt than to the proteins. Thus, weak ionic proteins are bound weakly