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Investigation of viral vectors using...

Negishi, Atsuko.

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Investigation of viral vectors using atomic force microscopy and microfluidic devices.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Investigation of viral vectors using atomic force microscopy and microfluidic devices./
Author:	Negishi, Atsuko.
Description:	127 p.
Notes:	Source: Dissertation Abstracts International, Volume: 65-12, Section: B, page: 6244.
Contained By:	Dissertation Abstracts International65-12B.
Subject:	Biophysics, Medical. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3156187
ISBN:	0496169823

Investigation of viral vectors using atomic force microscopy and microfluidic devices.
Negishi, Atsuko.

Investigation of viral vectors using atomic force microscopy and microfluidic devices. - 127 p.

Source: Dissertation Abstracts International, Volume: 65-12, Section: B, page: 6244.

Thesis (Ph.D.)--The University of North Carolina at Chapel Hill, 2004.

Researchers are modifying viruses into gene delivery vehicles in hope to cure diseases such as muscular dystrophy, hemophilia and cancer. Significant progress has been made toward this end, but further development and success of viral vectors depend on a deeper understanding of viral structure and physiology. Recent advances in microscopy have allowed new approaches to studying viruses that complement existing methodologies. Presented in this dissertation are novel viral studies using the atomic force microscope (AFM), a microscope that provides topographic information at the nanometer scale. As well microfluidic channels were used to study the effect of fluid flow properties on infection. A number of viruses are currently under study as potential vectors. We focus our studies on the adenovirus (Ad) and the adeno-associated virus (AAV) which have numerous attractive properties as vectors. The AFM is used to probe first, the structural aspects of the Ad and second, the virus-receptor interactions between AAV and its cell surface receptor, heparan sulfate proteoglycan (HSPG). The AFM was capable of imaging the capsid facets of intact Ad and DNA strands released from disrupted Ad capsids. In addition, we found that the stability of the capsid depended on the surface chemistry. An AFM-based binding assay was developed to study the binding between AAV and HSPG. The advantage of using the AFM for this purpose is its ability to simultaneously provide structural and quantitative information at the single molecule level. We measured a binding constant of 3.4 +/- 0.3 nM which is consistent with published reports. Microfluidic devices were used to study the dependence of fluid flow on infection. Cells were cultured in microfluidic channels and exposed to AAV vectors at various shear stresses. We found that a lower percentage of the cells were infected at higher shear stress. We also found that fluid forces can indirectly play a role in viral infection by influencing the cell state. A significantly lower percentage of cells that were treated with shear stress prior to vector exposure were infected compared to cells which were not exposed to shear stress.

ISBN: 0496169823Subjects--Topical Terms:

1017681
Biophysics, Medical.

Investigation of viral vectors using atomic force microscopy and microfluidic devices.
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500 $a Source: Dissertation Abstracts International, Volume: 65-12, Section: B, page: 6244.
500 $a Adviser: Richard Superfine.
502 $a Thesis (Ph.D.)--The University of North Carolina at Chapel Hill, 2004.
520 $a Researchers are modifying viruses into gene delivery vehicles in hope to cure diseases such as muscular dystrophy, hemophilia and cancer. Significant progress has been made toward this end, but further development and success of viral vectors depend on a deeper understanding of viral structure and physiology. Recent advances in microscopy have allowed new approaches to studying viruses that complement existing methodologies. Presented in this dissertation are novel viral studies using the atomic force microscope (AFM), a microscope that provides topographic information at the nanometer scale. As well microfluidic channels were used to study the effect of fluid flow properties on infection. A number of viruses are currently under study as potential vectors. We focus our studies on the adenovirus (Ad) and the adeno-associated virus (AAV) which have numerous attractive properties as vectors. The AFM is used to probe first, the structural aspects of the Ad and second, the virus-receptor interactions between AAV and its cell surface receptor, heparan sulfate proteoglycan (HSPG). The AFM was capable of imaging the capsid facets of intact Ad and DNA strands released from disrupted Ad capsids. In addition, we found that the stability of the capsid depended on the surface chemistry. An AFM-based binding assay was developed to study the binding between AAV and HSPG. The advantage of using the AFM for this purpose is its ability to simultaneously provide structural and quantitative information at the single molecule level. We measured a binding constant of 3.4 +/- 0.3 nM which is consistent with published reports. Microfluidic devices were used to study the dependence of fluid flow on infection. Cells were cultured in microfluidic channels and exposed to AAV vectors at various shear stresses. We found that a lower percentage of the cells were infected at higher shear stress. We also found that fluid forces can indirectly play a role in viral infection by influencing the cell state. A significantly lower percentage of cells that were treated with shear stress prior to vector exposure were infected compared to cells which were not exposed to shear stress.
590 $a School code: 0153.
650 4 $a Biophysics, Medical. $3 1017681
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