DEVELOPMENT AND CHARACTERIZATION OF HISTIDINE-TAGGED HPV16 L2 AND MS2-ARGININE-TAGGED RECOMBINANT PROTEINS FOR DOWNSTREAM PROCESSES
Date of Award
Open Access Master's Thesis
Master of Science in Biological Sciences (MS)
Administrative Home Department
Department of Biological Sciences
Committee Member 1
Committee Member 2
Human papillomaviruses (HPVs) are the most common sexually transmitted infections. Persistent infection with HPV can lead to anogenital cancers including head and neck cancers. Three prophylactic vaccines have been approved to prevent against some types of HPV infection. However, the vaccines are HPV-type specific and protect mostly against the HPV types included in the vaccines. To offer broader protection against more HPV types, studies in the field are developing candidate vaccines targeting a conserved minor capsid protein, L2. Nevertheless, reagents for developing and assessing L2 vaccines are limited. For example, antibodies to assess the antigenicity of some L2 epitopes are not available commercially and multivalent platforms to develop and purify clinical grade L2 antigens are limited. In this study, I developed and characterized the immunogenicity of a recombinant Histidine-tagged HPV16 L2 (amino acid 1-130) antigen. In addition to this, I explored the development of a multivalent display platform, a recombinant MS2-bacteriophage coat protein, with a C-terminal Arginine-tag for downstream purification using cation exchange chromatography. All Recombinant proteins (Histidine-tagged L2 and MS2-Arg tagged) were successfully expressed in bacteria. However, only Histidine-tagged L2 proteins were successfully purified in large quantity to homogeneity. Mice immunized with the Histidine-tagged L2 protein elicited anti-L2 IgG antibody titers greater than 103. The anti-L2 antibodies generated in this study will be valuable to researchers, in the field, developing L2 vaccines.
Alothaim, Tahiyat, "DEVELOPMENT AND CHARACTERIZATION OF HISTIDINE-TAGGED HPV16 L2 AND MS2-ARGININE-TAGGED RECOMBINANT PROTEINS FOR DOWNSTREAM PROCESSES", Open Access Master's Thesis, Michigan Technological University, 2017.
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