Masters Thesis

Separation of RNA and peptide using high performance liquid chromatography

High performance liquid chromatography (HPLC) has been widely used as an analytical tool for chemical and biochemical products. HPLC is derived in several techniques such as reverse phase chromatography, ion-exchange chromatography, and affinity chromatography. It depends on the nature of the analyte that determines what type of chromatography should be used. Moreover, there is not a universal method that can give the best resolution for every analyte. In this thesis, a method is developed to analyze several specific bio-products which have been shown to carry important information about the replication process in viruses. These are CPNT—an amino acid sequence at the N terminus of the coat protein, SL13 and Bbox—two RNA sequences responsible for interaction with the coat protein. These three sequences are found in Brome Mosaic Virus (BMV). They are crucial in the transcription and replication of a virus in the host cell when CPNT is bound to SL13 or Bbox. Therefore, investigating the interaction between them can give a better understanding about their mechanisms. An HPLC method was developed in this thesis to help analyze these sequences and help recycle the CPNT which will lower the cost of the research. We finally found that a gradient running for 30 minutes from 5% to 95% of solvent B with a flow rate of 1mL/min is a good condition to analyze these sequences by reverse phase chromatography. The solvent B is 0.1% TFA in acetonitrile and the solvent A is 0.1% TFA in water. The results also suggest a weak interaction between RNA and peptide complex since a complex peak did not show up in the chromatogram.

Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.