To browse Academia. Skip to main content. Log In Sign Up. Papers People. Production and purification of recombinant human granulocyte—macrophage colony stimulating factor GM-CSF from high cell density cultures of Pichia pastoris.
Gel Electrophoresis is a technique in molecular biology and biochemistry, which is used for the separation of proteins and nucleic acids, based on the motion of charged biological macromolecules in a constant electric field. Polyacrylamide gel separation is due to differences in molecular charge and separated differences of molecular weights and the configuration of molecules. There are so-called non-denaturating or native PAGE electrophoresis where the shared biological macromolecules are in the native state during electrophoresis and denaturing polyacrylamide gel electrophoresis where the samples were denatured beforehand in the case of nucleic acids using short heating the sample with glyoxal or formamide, to denature the proteins normally boiling the samples used in buffer containing a strong ionic detergent usually sodium dodecyl sulphate and the agent that disrupts the quaternary structure of the protein by breaking disulfide bridges between protein globules inside polypeptide chain — beta-mercaptoethanol. During the process of denaturing by PAGE electrophoresis, molecules are stored in the denatured state by the presence of chaotropic agents in the gel typically urea in the case of the PAGE electrophoresis of nucleic acids and proteins through the presence of the ion e.
Proteins are composed of aminic acids. All aminic acids are amphiprotic molecules dwelling of three types of amino acids: impersonal, acidic, and basic. Therefore, for any protein there is a characteristic pH, called the isoelectric point pi , at which the protein has no net charge and hence will non travel in the electric field.
This chapter focuses on the principle of electrophoresis and its utilization in a clinical laboratory. A sincere attempt has been made to discuss about clinical applications of serum protein electrophoresis, throwing light on the significance of serum protein electrophoresis in the management of multiple myeloma. Emphasis has been made on quality assurance in terms of accuracy and precision in electrophoresis to ensure reliability of patient results. A note on issues with lack of standardization of reporting of electrophoresis and an insight into global efforts to standardize the reporting of the assay has been included in this chapter.