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- Member of: Theses and Dissertations
- Resource Type: Text
- Status: Published
drugs in basic research, biotechnology, diagnostics and therapeutics. However, due to the
cost, labor and time associated with production of antibodies focus has recently changed
towards potential of peptides to act as protein affinity reagents. Affinity peptides are easy
to work with, non-immunogenic, cost effective and amenable to scale up. Even though
researchers have developed several affinity peptides, we are far from compiling library of
peptides that encompasses entire human proteome. My thesis describes high throughput
pipeline that can be used to develop and characterize affinity peptides that bind several
discrete sites on target proteins.
Chapter 2 describes optimization of cell-free protein expression using commercially
available translation systems and well-known leader sequences. Presence of internal
ribosome entry site upstream of coding region allows maximal expression in HeLa cell
lysate whereas translation enhancing elements are best suited for expression in rabbit
reticulocyte lysate and wheat germ extract. Use of optimal vector and cell lysate
combination ensures maximum protein expression of DNA libraries.
Chapter 3 describes mRNA display selection methodology for developing affinity peptides
for target proteins using large diversity DNA libraries. I demonstrate that mild denaturant
is not sufficient to increase selection pressure for up to three rounds of selection and
increasing number of selection rounds increases probability of finding affinity peptide s.
These studies enhance fundamental understanding of mRNA display and pave the way
for future optimizations to accelerate convergence of in vitro selections.
Chapter 4 describes a high throughput double membrane dot blot system to rapidly
screen, identify and characterize affinity peptides obtained from selection output. I used
dot blot to screen potential affinity peptides from large diversity of previously
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uncharacterized mRNA display selection output. Further characterization of potential
peptides allowed determination of several high affinity peptides from having Kd range 150-
450 nM. Double membrane dot blot is automation amenable, easy and affordable solution
for analyzing selection output and characterizing peptides without ne ed for much
instrumentation.
Together these projects serve as guideline for evolution of cost effective high throughput
pipeline for identification and characterization of affinity peptides.
This thesis attempts to achieve the research objectives by examining the LEED certified buildings on the Arizona State University (ASU) campus in Tempe, AZ, from two complementary perspectives: the Macro-level and the Micro-level. Heating, cooling, and electricity data were collected from the LEED-certified buildings on campus, and their energy use intensity was calculated in order to investigate the buildings' actual energy performance. Additionally, IEQ occupant satisfaction surveys were used to investigate users' satisfaction with the space layout, space furniture, thermal comfort, indoor air quality, lighting level, acoustic quality, water efficiency, cleanliness and maintenance of the facilities they occupy.
From a Macro-level perspective, the results suggest ASU LEED buildings consume less energy than regional counterparts, and exhibit higher occupant satisfaction than national counterparts. The occupant satisfaction results are in line with the literature on LEED buildings, whereas the energy results contribute to the inconclusive body of knowledge on energy performance improvements linked to LEED certification. From a Micro-level perspective, data analysis suggest an inconsistency between the LEED points earned for the Energy & Atmosphere and IEQ categories, on one hand, and the respective levels of energy consumption and occupant satisfaction on the other hand. Accordingly, this study showcases the variation in the performance results when approached from different perspectives. This contribution highlights the need to consider the Macro-level and Micro-level assessments in tandem, and assess LEED building performance from these two distinct but complementary perspectives in order to develop a more comprehensive understanding of the actual building performance.
This dissertation fills the crucial knowledge gap in contract administration functions and tools for DB and CM/GC highway project delivery. First, this research identifies and models contract administration functions in DBB, CM/GC, and DB using integrated definition modeling (IDEF0). Second, this research identifies and analyzes DB and CM/GC tools for contract administration by conducting 30 ACM project case studies involving over 90 ACM practitioners. Recommendations on appropriate use regarding project phase, complexity, and size were gathered from 16 ACM practitioners. Third, the alternative technical concepts tool was studied. Data from 30 DB projects was analyzed to explore the timing of DB procurement and DB initial award performance in relation to the project influence curve. Types of innovations derived from ATCs are discussed. Considerable industry input at multiple stages grounds this research in professional practice.
Results indicate that the involvement of the contractor during the design phase for both DB and CM/GC delivery creates unique contract administration functions that need unique tools. Thirty-six DB and CM/GC tools for contract administration are identified with recommendations for effective implementation. While strong initial award performance is achievable in DB projects, initial award performance in this sample of projects is only loosely tied to the level of percent base design at procurement. Cost savings typically come from multiple ATCs, and innovations tend to be incremental rather than systemic, disruptive, or radical. Opportunity for innovation on DB highway projects is influenced by project characteristics and engaging the DB entity after pre-project planning.
The built environment is responsible for a significant portion of global waste generation.
Construction and demolition (C&D) waste requires significant landfill areas and costs
billions of dollars. New business models that reduce this waste may prove to be financially
beneficial and generally more sustainable. One such model is referred to as the “Circular
Economy” (CE), which promotes the efficient use of materials to minimize waste
generation and raw material consumption. CE is achieved by maximizing the life of
materials and components and by reclaiming the typically wasted value at the end of their
life. This thesis identifies the potential opportunities for using CE in the built environment.
It first calculates the magnitude of C&D waste and its main streams, highlights the top
C&D materials based on weight and value using data from various regions, identifies the
top C&D materials’ current recycling and reuse rates, and finally estimates a potential
financial benefit of $3.7 billion from redirecting C&D waste using the CE concept in the
United States.