Chemistry and Biochemistry Copyright (c) 2013 California Polytechnic State University All rights reserved. http://digitalcommons.calpoly.edu/chemsp Recent documents in Chemistry and Biochemistry en-us Fri, 25 Jan 2013 16:53:47 PST 3600 Design and Experimental Validation of Continuous DNA Separation using ITP-Based Microfluidic Device http://digitalcommons.calpoly.edu/chemsp/6 http://digitalcommons.calpoly.edu/chemsp/6 Thu, 19 Jan 2012 17:15:10 PST We have demonstrated a chip-based microfluidic device suitable for rapid and high-throughput DNA separation and purification for downstream analysis. The glass-based device incorporates a transverse free-flow isotachophoresis (tFF-ITP) that overcomes the volume limitations of CE approaches, and allows continuous sample processing in contrast to batch-mode solid-phase extraction (SPE). The device is able to focus DNA at flow rates up to 100 uL/min, and sample conductivity up to 2 mS/cm. Downstream of the chip, 30-40% of DNA from the input sample is recovered as a result of ITP focusing, and preliminary results indicate that this design is able to purify DNA from contaminating species, particularly those that inhibit PCR.

]]> Nora C. M. Goscinski et al. SYNTHESIS OF AN ANTIMICROBIAL TEXTILE COATING http://digitalcommons.calpoly.edu/chemsp/5 http://digitalcommons.calpoly.edu/chemsp/5 Tue, 10 Jan 2012 17:32:21 PST A titania nanosol was synthesized and coated onto nylon/cotton blended textile substrates. The substrates were characterized via SEM for adhesion and nanoparticle formation, then subjected to antimicrobial efficacy tests. The titania nanosol was successfully coated on to textiles samples. Particles were observed to be around 2 by 3 micrometers and formed between the interstitial space of textile fibers. Although larger than typical nanoparticles, the coatings exhibited what seemed to be antimicrobial activity. Titania nanosol coated textile samples were subjected to Kirby Bauer Assay in the presence of S. aureus. The coated textile sample exhibited an inhibition of growth around its edges while the uncoated sample encouraged growth. A post-antibiotic effect was observed to be 1.2 hours on S. aureus when exposed to the titania coated textile.

]]> William M. Morris LIFE AS A SWIMMER http://digitalcommons.calpoly.edu/chemsp/4 http://digitalcommons.calpoly.edu/chemsp/4 Wed, 23 Nov 2011 09:54:37 PST Danielle N. Coville Characterization of Amino Acid Residues Integral to Neuronal Binding of Amyloid Beta Protein in Alzheimer’s Disease http://digitalcommons.calpoly.edu/chemsp/3 http://digitalcommons.calpoly.edu/chemsp/3 Tue, 05 Apr 2011 16:34:19 PDT Purpose: Alzheimer’s Disease is a neurodegenerative disease resulting from over-production and neuronal accumulation of amyloid-beta proteins (Aβ40/Aβ42). The glycine residue at position 33 and histidine residues at positions 13 and 14 are involved with binding and internalization of these proteins, actions potentially inhibited by substituting or sterically hindering these residues with an antibody specific to positions 2-11 (IgG-4.1). Rat pheochromocytoma (PC12) cells differentiated with nerve growth factor were used as a neuronal model to determine whether substitution and/or antibody block amyloid-beta’s neuronal interactions.

Methods: PC12 cells were incubated with fluorescein-labeled-amyloid-beta-40 (F-Aβ40) or substituted F-Aβ40 derivatives (F-Aβ40-H13,14G, F-Aβ40-H13,14G;G33A), with or without IgG-4.1. Cells were analyzed by flow cytometry.

Results: PC12 cells incubated with F-Aβ40 and IgG-4.1, as well as cells incubated with substituted peptides, exhibit decreased mean fluorescence compared to cells incubated with F-Aβ40 alone.

Conclusion: IgG-4.1 and amino acid substitution decreased binding and internalization of Aβ40, suggesting immunotherapy may reduce intraneuronal accumulation of amyloid-beta.

]]> Nicole C. Olson OVEREXPRESSION AND PARTIAL PURIFICATION OF RECOMBINANT HUMAN SERINE HYDROXYMETHYLTRANSFERASE: A POTENTIAL ANTICANCER TARGET http://digitalcommons.calpoly.edu/chemsp/2 http://digitalcommons.calpoly.edu/chemsp/2 Wed, 02 Jun 2010 10:06:02 PDT Serine Hydroxymethyltransferase (SHMT) is a member of the thymidylate synthase cycle that provides pyrimidine nucleotides to actively dividing cells, including cancer cells. The other two enzymes involved in this cycle have been targets for clinical anticancer drugs, suggesting that inhibitors of SHMT may also be used as clinical anticancer drugs. In order to test potential inhibitors against SHMT, human SHMT has been overexpressed in E. coli and partially purified by selective ammonium sulfate fractionation followed by anion-exchange fast protein liquid chromatography. A coupled assay using L-allo-threonine and alcohol dehydrogenase was used to identify active fractions, which were analyzed for purity by SDS-PAGE. The affinity purification of human SHMT by Ni+2-NTA affinity chromatography trials concluded that it was not expressed as a fusion protein containing a polyhistidine affinity tag. The gene encoding for human SHMT was then cloned into a pET151/D-TOPO (Invitrogen) plasmid for better purification. A fraction of transformed colonies were screened for the cloned gene by PCR amplification and restriction enzyme digestion of isolated plasmid DNA. None of the colonies screened were found to contain the full-length hSHMT gene.

]]> Suprit Singh Deol Overexpression and Partial Purification of Serine Hydroxymethyltransferase http://digitalcommons.calpoly.edu/chemsp/1 http://digitalcommons.calpoly.edu/chemsp/1 Mon, 14 Dec 2009 11:53:25 PST Kimberly Rose Stevens