Montana Kaimin, January 18, 2024

Student newspaper of the University of Montana, Missoula.https://scholarworks.umt.edu/studentnewspaper/11135/thumbnail.jp

HPLC Column Packing Efficiency: For Halogen Bonding Separation Techniques

High-performance liquid chromatography (HPLC) is an important technique for the separation and analysis of mixtures of nonvolatile compounds such as proteins, peptides, and pharmaceuticals. Stainless steel tubes (“columns”) uniformly packed with chemically modified small-diameter spherical porous silica particles are essential to the method. As part of the “Halogen Bonding Liquid Chromatography” laboratory research with Dr. Christopher Palmer and Dr. Orion Berryman, silica particles are modified with unique chemistries to further develop HPLC separation capabilities. These particles, generated in relatively small quantities, must be packed into columns to allow evaluation of their performance. The halogen bonding modified silica project aims to pack columns with 200-300mg of modified silica. Although thousands of commercial HPLC columns are packed and sold yearly, effective packing techniques and methods are generally proprietary. Published studies (Wahab 2017) suggest using a silica slurry that is 5 to 8% silica concentration to produce quality HPLC columns. In the current study, we have developed and optimized methods utilizing minimum quantities of silica, either through lower slurry concentrations or reduced slurry volumes, to pack high-performance columns. Unmodified spherical silica was used to develop an effective column-packing method utilizing minimum quantities of silica. A series of columns were packed at different silica concentrations and under different packing conditions. Various techniques were investigated to enable a higher-quality column using low slurry concentrations. The packing pressure and solvent used during the packing process are two characteristics of column packing that are optimized for a low silica concentration column to be packed correctly. Being packed properly ensures that silica is packed uniformly throughout the column without any spaces or bubbles in the column itself. The quality of each column is evaluated through specific HPLC methods to measure peak width, shape, and retention time, with optimally packed columns producing narrow and symmetrical peaks. The method\u27s flow rate, oven temperature, and injection volume are varied to allow for the best performance. The final step is to show the method\u27s reproducibility by repacking and testing under the same conditions. Producing an efficient low silica concentration HPLC column is crucial to the research team\u27s success and will allow a halogen-boding liquid chromatography column to be constructed and commercialized. Citations Wahab, M. F., Patel, D. C., Wimalasinghe, R. M., & Armstrong, D. W. (2017). Fundamental and practical insights on the packing of modern high-efficiency analytical and capillary columns. Analytical Chemistry, 89(16), 8177–8191. https://doi.org/10.1021/acs.analchem.7b0093

PRDM3 Protein Purification: An Optimized Approach for Structural Understanding

Investigating proteins involved in driving cancer progression is crucial for advancing therapeutic strategies. PRDM3 (PR/SET domain-containing 3) has emerged as a key player in sustaining ovarian tumors. To facilitate an in-depth study of PRDM3, obtaining the protein in substantial quantities with a high-level purity through recombinant expression is imperative. The methodology comprises a series of steps: integrating the PRDM3 gene fused with an affinity tag into a plasmid; transferring the engineered plasmid into bacterial cells; expressing the affinity-tagged protein (fusion protein) within the bacterial cells and harvesting them. Cell lysis and protein extraction are then conducted, followed by affinity chromatography for fusion protein purification and subsequent cleavage of the affinity tag. Gel filtration chromatography is used for additional purification, resulting in a homogeneous PRDM3 protein for detailed structural analyses. Achieving maximum purity and a high yield in the purification of PRDM3 poses challenges in protein extraction, tag cleavage and homogeneity attainment. This can significantly impact the feasibility of subsequent protein studies, necessitating optimization strategies. This study aims to address these challenges by refining the protein purification methods for PRDM3, facilitating a more precise understanding of its interactions with other proteins and its role in oncogenic processes. Standard chromatographic separations, such as affinity and gel filtration chromatography, combined with an affinity tag and the use of detergent, have been utilized to optimize protein extraction. Simultaneously, changes in cleaving conditions have been introduced to minimize protein precipitation post-tag cleavage. The introduction of these combined enhancements distinguishes this study from conventional methods and demonstrates a novel strategy for optimizing PRDM3 protein extraction and purification, ultimately ensuring its isolation with optimal purity and yield. In conclusion, this study lays the groundwork for more in-depth investigations into the molecular interplay between PRDM3 and other proteins- potentially leading to advances in therapeutics for ovarian cancer

SB19-23/24: Resolution Encouraging Excused Student Absences for the Democracy Summit

This resolution passed 16Y-0N-1A on a roll call vote during the March 6, 2024 meeting of the Associated Students of the University of Montana (ASUM)

Deeply Rooted: The Role of Indigenized Research Approaches in Camas Restoration Ecology

Despite being one of the most culturally significant and historically abundant native plants of the Pacific Northwest, common camas (Camassia quamash (Pursh) Greene) has undergone a precipitous decline across its range. This decline, driven primarily by agricultural land-use conversion and wetland drainage, has disproportionately impacted Indigenous peoples for whom camas is a cultural keystone species. For thousands of years prior to colonization, tribes across the Pacific Northwest relied on camas bulbs as a first food and managed camas prairies using Traditional Resource Management (TRM) practices. Camas restoration has emerged as a widespread management priority in recent years, particularly among Indigenous groups, and many of the recent studies on the subject have addressed the species’ cultural significance or drawn upon Traditional Ecological Knowledge (TEK). TEK, which refers to the intimate knowledge of local ecosystems held by Indigenous peoples, has been widely embraced in the field of restoration ecology as a complementary epistemology to western science, though serious concerns have emerged regarding the ethical dimensions of its use in academia. In response to these concerns, a growing number of studies in the field of restoration ecology have begun to utilize Two-Eyed Seeing and other research approaches to respectfully and meaningfully integrate the perspectives, priorities, and values of the Indigenous communities impacted by both ecological degradation and restoration. I review the current state of camas restoration ecology, outline a number of Indigenized research approaches both within the camas literature and beyond, and highlight how these approaches stand to improve the social and ecological outcomes of camas restoration. I then outline several open questions in camas restoration ecology that could be addressed using these approaches, primarily concerning the mechanisms by which traditional management practices increase camas abundance

Poster Session #1 and Franke GLI Presentations

Posters South Ballroom: Poster Session #1 GLI Presentations UC 333: Franke GLI Presentations (10:45-12:15

UMCUR 2024 Program PDF

This program includes the schedule for oral presentations and posters