Phage Display Techniques for Selection of Inorganic Binding Peptides with Electroactive Properties for Biosensor Applications

MISSING IMAGE

Material Information

Title:
Phage Display Techniques for Selection of Inorganic Binding Peptides with Electroactive Properties for Biosensor Applications
Physical Description:
1 online resource (167 p.)
Language:
english
Creator:
Yeh, Ya-Wen
Publisher:
University of Florida
Place of Publication:
Gainesville, Fla.
Publication Date:

Thesis/Dissertation Information

Degree:
Doctorate ( Ph.D.)
Degree Grantor:
University of Florida
Degree Disciplines:
Biomedical Engineering
Committee Chair:
GOWER,LAURIE B
Committee Co-Chair:
ORMEROD,BRANDI K
Committee Members:
TSENG,YIIDER
GULIG,PAUL A

Subjects

Subjects / Keywords:
biosensor
Biomedical Engineering -- Dissertations, Academic -- UF
Genre:
Biomedical Engineering thesis, Ph.D.
bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract:
In biosensor devices, functionalization of the surface with covalent linkers is usually employed. However, it is difficult to avoid the loss of activity following the bioreceptor-analyte binding event, which limits the lifetime of the device. The goal of this project was to use phage display to biopan for inorganic binding peptides that are reversible upon application of an electric field. This technique could provide dynamic functionalization of surfaces, with applications such as reconfigurable and self-cleaning devices.  In this study, biopanning was performed to select for phage with displayed peptides that bind strongly to indium zinc oxide (IZO), a transparent semiconducting oxide which is an attractive electrode for biosensor applications. IZO binding phage collected by two different chemical elution methods, low pH or high salt elution buffer, were identified. It was found that phage clones eluted with high salt buffer were more selective in their binding to IZO than the clones eluted by low pH buffer. An electro- releasing device was then developed to select from those clones the ones that could be released upon application of an electric field.  In an alternative method, because a strong binding peptide may not be a reversible peptide, a new biopanning protocol was developed with an electro-elution process instead of the regular chemical elution. Using a small electronic device in the biopanning media, phage that desorb upon application of a field could be collected directly. Clones selected by electro-elution displayed a different composition of amino acids as compared to those from the chemical elution approach.  Based on the studies of phage clones selected from chemical elution and electro- elution biopanning processes, the amino acid sequences of the displayed peptides were then synthesized to further test the binding and release properties of the peptides in isolated form.  The binding affinity of the synthetic IZO binding peptides was confirmed. It was found that both the charge character and the secondary structure of a peptide seemed to play a role in affecting the electroactive properties. In conclusion, my work demonstrates that phage display with a novel electro-elution step can select peptides that are more sensitive to an electric field.
General Note:
In the series University of Florida Digital Collections.
General Note:
Includes vita.
Bibliography:
Includes bibliographical references.
Source of Description:
Description based on online resource; title from PDF title page.
Source of Description:
This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Statement of Responsibility:
by Ya-Wen Yeh.
Thesis:
Thesis (Ph.D.)--University of Florida, 2013.
Local:
Adviser: GOWER,LAURIE B.
Local:
Co-adviser: ORMEROD,BRANDI K.
Electronic Access:
RESTRICTED TO UF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE UNTIL 2015-12-31

Record Information

Source Institution:
UFRGP
Rights Management:
Applicable rights reserved.
Classification:
lcc - LD1780 2013
System ID:
UFE0046200:00001