Browsing by Subject "Serum albumin"
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Item Development of cross-reactive receptors based on serum albumin(2015-05) Diehl, Katharine Louise; Anslyn, Eric V., 1960-; Brodbelt, Jennifer S; Iverson, Brent L; Kerwin, Sean M; Sessler, Jonathan LIn recent years, differential sensing has become an increasingly popular approach to molecular recognition. Mimicking the mammalian senses of taste and smell, arrays of semi-selective sensors generate a fingerprint for each analyte. Pattern recognition algorithms allow these arrays to be used for discriminating analytes and for predicting the identity of unknowns. Arrays of cross-reactive receptors have found use in a variety of sensing applications, including the differentiation of biologically-relevant analytes (Chapter 1). Serum albumin has previously been used in an array format for the discrimination of hydrophobic analytes such as terpenes, plasticizers, and fatty acids. This protein is a versatile cross-reactive receptor because of its ability to bind hydrophobic analytes with different affinities and in different modes. In Chapter 2, the use of serum albumin to pattern hydrophobic analytes was further expanded to include glycerides. Glycerides are challenging analytes because they are structurally similar to one another. Due to difficulties in identifying the regio- and stereochemistry of the unsaturated glycerides, a sample pretreatment consisting of olefin cross metathesis was used prior to array analysis. Using the array, twenty glycerides were discriminated, including stereo- and regioisomeric pairs. Further, glycerides in mixtures were quantitated. Due to the success with using serum albumins as receptors for hydrophobic analytes, it was hypothesized that serum albumin could be used in a different way to develop receptors for other types of analytes. The serum albumin functions as a scaffold onto which species dynamically assemble through hydrophobic interactions. In Chapter 3, the use of fatty acid-appended recognition units was explored as a way of building such receptors. While these conjugates did not bind to the protein as well as expected, an alternative strategy was explored in Chapter 4 with squaraines and thiols. As a starting point toward this goal, a series of squaraines were synthesized, and their reactivity to thiols and affinity for serum albumin were investigated. Finally, in Chapter 5, the reversible covalent attachment of recognition units to serum albumin with thia-Michael chemistry was explored. A bifunctional conjugate acceptor was used to reversibly label proteins with thiols as well as to generate resin-bound dynamic combinatorial libraries.Item Differential sensing of hydrophobic analytes with serum albumins(2012-05) Ivy, Michelle Adams; Anslyn, Eric V., 1960-In the last decade, there has been a growing interest in the use of differential sensing for molecular recognition. Inspired by the mammalian olfactory system, differential sensing employs an array of non-selective receptors, which through cross-reactive interactions, create a distinct pattern for each analyte tested. The unique fingerprints obtained for each analyte with differential sensing are studied with statistical analysis techniques, such as principal component analysis and linear discriminant analysis. It was postulated that serum albumin proteins would be applicable to differential sensing schemes due to significant differences in sequence identity between different serum albumin species, and due to the wide range of hydrophobic molecules which are known to bind to these proteins. Consequently, cross-reactive serum albumin arrays were developed, utilizing hydrophobic fluorescent indicators to detect hydrophobic molecules. As such, serum albumin cross-reactive arrays were employed to discriminate subtly different hydrophobic analytes, and mixtures of these analytes, in the form of terpenes and perfumes, plasticizers and plastic explosive mixtures, and glycerides and adipocyte extracts. In this doctoral work, a detailed review of the field of differential sensing, and a thorough study of principal component analysis and linear discriminant analysis in various differential sensing scenarios, are given. These introductory chapters aid in better understanding the methods and techniques applied in later experimental chapters. In chapter 3, serum albumins, a PRODAN indicator, and an additive are shown to discriminate five terpene analytes and terpene doped perfumes. Chapter 4 describes an array with serum albumins, two dansyl fluorophores, and an additive which successfully differentiate the plasticizers found within the plastic explosives C4 and Semtex and simulated C4 and Semtex mixtures. Discrimination of these simulated mixtures was also achieved with this array in the presence of soil contaminants, demonstrating the potential real-world applicability of this sensing ensemble. Finally, chapter 5 details an array consisting of serum albumins, several fluorescent indicators, and a Grubb's olefin metathesis reaction, to differentiate saturated and unsaturated triglycerides, diglycerides, and monoglycerides. Mixtures of glycerides in adipocyte extracts taken from rats with different health states were then successfully discriminated, showing promise for clinical applications in differentiating adipoctyes from pre-diabetic, type 2 diabetic, and non-diabetic individuals.Item The primary binding site of pyridoxol 5'-phosphate to bovine serum albumin(Texas Tech University, 1970-08) Grandjean, Carter JulesNot available