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Purification of Phage-Displayed HSA-Specific Peptide for Biosensor Production

Huber, Alexander Domenico
http://rave.ohiolink.edu/etdc/view?acc_num=ysu1559730074086289

Abstract Details

2019, Master of Science in Biological Sciences, Youngstown State University, Department of Biological Sciences and Chemistry.
The goal of this project is to produce a carbon nanotube transistor-biosensor that can detect blood, which can be used by the police or military. This biosensor would be able to detect human serum albumin (HSA) and send a distress signal for help. HSA is the most abundant component of blood plasma, making it an ideal target for detection. The specific goal of this study is to purify an HSA-specific heptapeptide, BR-1, from a M13 filamentous phage that was obtained through phage display. The specificity of BR-1 for HSA was confirmed by performing an Enzyme-linked Immunosorbent Assay (ELISA), however, several peptide ELISAs were performed beforehand in order to find the optimal conditions for the ELISA (biotin amounts on peptide for streptavidin-biotin system and wash and blocking buffers). The BR-1 peptide was previously sequenced in this lab, and this sequence was utilized for this experiment. Primers were designed for a polymerase chain reaction process that incorporated the peptide sequence into the expression vector pMal-c5x. This DNA was then subsequently purified, ligated, and sequenced again to confirm the presence of the peptide sequence in the vector. The majority of the peptide sequence was incorporated into the vector; however, this process will need to be repeated in order to incorporate the complete sequence into the vector. After the peptide sequence is incorporated into the pMal-c5x expression vector, this vector will then be transformed into E. coli, and will be induced to express this peptide fused to a maltose-binding protein (MBP). The peptide will subsequently be purified from the MBP using an amylose resin column. After this process is complete, the specificity of this purified peptide can be tested by performing another ELISA. Future efforts will be to use the peptide that our lab purified and incorporate it into a biosensor that can detect HSA.
Diana Fagan, PhD (Advisor)
David Asch, PhD (Committee Member)
Jonathan Caguiat, PhD (Committee Member)
81 p.
Biology
Biosensor; Human serum albumin; Phage display; pMal-c5x

Recommended Citations

Citations

  • Huber, A. D. (2019). Purification of Phage-Displayed HSA-Specific Peptide for Biosensor Production [Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1559730074086289

    APA Style (7th edition)

  • Huber, Alexander. Purification of Phage-Displayed HSA-Specific Peptide for Biosensor Production. 2019. Youngstown State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1559730074086289.

    MLA Style (8th edition)

  • Huber, Alexander. "Purification of Phage-Displayed HSA-Specific Peptide for Biosensor Production." Master's thesis, Youngstown State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1559730074086289

    Chicago Manual of Style (17th edition)

ysu1559730074086289
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© 2019, all rights reserved.
This open access ETD is published by Youngstown State University and OhioLINK.