News and Events for the Department of Chemical and Biochemical Engineering in the College of Engineering and Information Technology at UMBC.
Aditi Bhaskar successfully defended her Ph.D. on November 12th, 2013. Her research was performed in the Welty Lab.
Shaunak Uplekar successfully defended his Ph.D. on November 12th, 2013. His dissertation was entitled "Impact of process parameters on product titer and quality attributes during cell culture scale down."
Date: Tuesday, Nov. 12th, 2013
Time: 12:00 pm
Location: TRC 206
Light refreshments will be served at 11:45 am
Dissertation title: Impact of process parameters on product titer and quality attributes during cell culture scale down
Quality by Design and Process Analytical Technology regulatory initiatives aim to achieve consistent production of biopharmaceutical products of predefined quality. Extensive process development studies are imperative to establish such a production process. Currently, these studies are primarily being carried out in bench scale systems. However, bench scale bioreactor studies are costly, tedious and time consuming to set up. An in-house developed, novel high-throughput minibioreactor system shows a significant potential to improve and streamline process development studies. However, in order to fully qualify this system as a scale down model, comparability between the two scales needs to be established. Comparability of product titers and product quality aspects such as glycosylation profile, which is considered to be one of the major product quality attributes, would be of greatest relevance.
Here, we investigate cell culture scale down on monoclonal antibody titers and their N-glycan profiles produced by serum free mammalian cell culture in these two systems. Methods for purification of monoclonal antibody using protein-A chromatography followed by ion exchange chromatography and N-glycan analysis using high pH anion exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) were developed. Structural determination of glycans was done using MALDI TOF mass spectrometry. A comparability experiment indicated that DO (dissolved oxygen) and pH profiles, cell growth, glucose and lactate profiles were similar in bench scale bioreactors and minibioreactors. Although the relative areas of major N-glycans obtained by HPAEC were found to be comparable in two systems, antibody titers in bench scale bioreactor were about 50% higher than in minibioreactor. While investigating this, a noticeable difference in the glutamine consumption in two systems was observed. Evidence in the literature related glutamine consumption to pCO2 levels. In order to investigate the role of pCO2, a novel pCO2 sensor patch was modified for use in mammalian cell culture. This sensor revealed a difference between the pCO2 profiles in both systems. CO2 stripping studies were conducted in order to have comparable pCO2 stripping rate. A comparability study conducted at comparable pCO2 stripping rate showed similar pCO2 profiles and a significant improvement in product titers in minibioreactors was achieved. Finally, the effect of dissolved oxygen (DO) in minibioreactors and bench scale bioreactors was investigated. Effects of DO on cell growth, viability, glucose and lactate profiles, product titers and their N-glycan profiles were found to be comparable. This project attempted to understand the changes that occur during scale down in the cell culture environment, that are significant enough to impact critical process outcomes such as product titer and product quality . This understanding proved to be crucial to provide the comparability of these miniaturized systems with the bench scale systems and to establish them as a scaled down model.
Mid-Atlantic Biosolids Association and the Department of Chemical, Biochemical, and Environmental Engineering present:
The Science of Biosolids: Current Research and Implications for Management
Tuesday, November 19th, 2013
University Center Ballroom
8:00 AM - 4:00 PM
For more information and to register, visit "http://www.mabiosolids.org/index.php?page=events-calendar"
Alba Torrents, Professor
Environmental Engineering Program
Civil and Environmental Engineering
University of Maryland, College Park
Date: November 11, 2013
Time: 12:00pm - 1:00pm
Location: TRC 206
Title: Re-Use of Wastewater Effluents: Benefits and Concerns
Through the natural water and nutrient cycle, the earth has recycled and reused water and nutrients for millions of years. Currently, 1.6 billion people live in countries and regions with water and food scarcity and, with population growth and climate change, the number is expected to rise to 2.8 billion people by 2025. Traditional urban wastewater systems need to be transformed in a sustainable, closed-loop urban wastewater management based in the conservation of water and nutrients. There is concern that along with organic matter, nutrients and micronutrients, biosolids could be a source of anthropogenic microconstituents. For over 15 years, our group has been developing methodologies to assess the fate of organic micro pollutants introduced in the environment. We know that many of such micro pollutants are endocrine disruptors and they make their way into the environment, yet many questions remain on their fate and potential risks. A combination of “in-pant”; controlled and field scale studies will be presented and discussed.
A recent article on the Smithsonian Science website highlights the work of CBEE professor Upal Ghosh, along with Cynthia Gilmour from the Smithsonian Environmental Research Center, and their colleagues. These researchers have found a new low-cost, nonhazardous way to reduce the toxicity of hot spots polluted with mercury by using charcoal to trap it in the soil.
The complete Smithsonian Science article can be found at this link and the study "Activated Carbon Mitigates Mercury and Methylmercury Bioavailability in Contaminated Sediments," published in the journal Environmental Science & Technology, can be found here
Christopher Kistler, Associate Principle Scientist for Merck
Date: Monday, October 21
Time: 12:00pm - 1:00pm
Location: TRC 206
Title: Technology Improvements to Accelerate Process Development of Biologics
Christopher Kistler, Krista Alvin, Rachel Bareither, and David Pollard
Biologics New and Enabling Technologies, Merck & Co., Inc., 126 E. Lincoln Ave, Rahway, NJ
Pressures continue to reduce the time from discovery to product launch and minimize the costs of not only manufacturing but also process development. This is particularly difficult for upstream development where large DOE designs in lab scale reactors, provides a significant equipment and resource constraint. The presentation will show how innovation through automation and single-use technology has led to more efficient process development. This includes the creation of multi use tools to handle both cell culture and microbial expression platforms. Improvements will be shown for end to end development focusing on upstream processing followed by purification and analytics. Examples include the use of cell line development automation for the elimination of manual shakeflask stages and increase throughput of clonal evaluation. For more advanced process development a novel small scale single use prototype bioreactor is evaluated. This system is designed for automated multi tank experimentation with robotic sampling, feeding and control. This is integrated with high throughput purification and analytics using a systematic approach of statistical design of experiments in combination with 'omics' technologies. This allows for a rapid end to end process development and builds a fundamental understanding of the impact of process operations to control process consistency and product quality.
Congratulations to Niloufar Pezeshk, who successfully defended her M.S. on September 20th, 2013. Her research was performed in the Good Lab; the title of her dissertation was Inhibiting Amyloid-beta fibrils from binding to cells
Date: Friday, August 16th, 2013
Time: 10:00 am
Location: ITE 456
Light refreshments will be served at 9:45 am
Dissertation title: Proteomic Analysis of Aspergillus nidulans during Autophagy and the Role of Autophagy Genes Anatg13 and Anatg8
Aspergilli represent an extremely important genus of microorganisms which can be both harmful pathogens, and beneficial pharmaceutical producers. In Aspergilli’s interactions with man, suboptimal nutrient conditions are often present, and lead to a phenomenon known as autophagy. Autophagy is a cellular recycling mechanism that (in the case of macroautophagy) is augmented under nutrient limited conditions to recycle cytoplasmic macromolecules and organelles for use in essential cell functions. Strategic manipulation of autophagy could ultimately lead to improved bioprocesses or anti-fungal treatments. Using the model filamentous fungus Aspergillus nidulans, a number of important questions about autophagy have been addressed.
Critical to the study of autophagy is the balance between self-degradation and self-preservation. Therefore, we adapted an XTT metabolic activity assay for use in filamentous fungi. The assay was first tested using a number of bioprocess related stresses (e.g. temperature, shear), and found to be superior to DCW as an assessment of culture health. Next, the metabolic activity of fungal cultures was tested during autophagy inducing conditions, demonstrating that the autophagy capable TN02A3 strain was more viable than an autophagy deficient ∆atg13 strain in nutrient limiting conditions.
By analyzing the proteome of key autophagy mutants ∆Anatg13 and ∆Anatg8, an improved molecular understanding of autophagy in filamentous fungi was achieved. Using 2-dimensional electrophoresis, 44 unique proteins were observed with significant expression changes caused either by addition of rapamycin (a chemical inducer of autophagy) or deletion of Anatg13. AnAtg13 dependent changes of multiple ribosomal proteins and a key polyamine biosynthetic protein, spermidine synthase (AnSpdA), provides molecular evidence of AnAtg13 dependent lifespan extension in A. nidulans.
After establishing improved shotgun proteomic methods on the Thermo LTQ-XL, we generated a more thorough assessment of the A. nidulans response to autophagy induction by measuring protein expression as a function of time. It was found that autophagy induction caused a rapid and sustained increase in proteolysis, amino acid degradation, and lipid metabolism. These changes occurred in both the control (TN02A3) and autophagy deficient ∆Anatg8 strains. Many of the proteins with a delayed change in expression were autophagy dependent. These include proteins involved in those involved in secretion, hydrolysis of alternative carbon sources, and secondary metabolite production; all of which are important to the bioprocess industry.
In January 2013, the EWB-UMBC team traveled to Isongo, Kenya for an assessment trip, during which they successfully established relationships with the residents, mapped the community, analyzed the water quality, and surveyed residents on water, sanitation, and hygiene. Now the group is planning an implementation trip for January 2014 to install a clean water system that will benefit 500 people from Isongo.
But they can’t make the trip without your help.
How can you help?
Your donation makes two things possible: (1) Development of a clean water supply for the community of Isongo, Kenya and (2) transformative experiences for UMBC students interested in international, humanitarian work. You can also help the EWB-UMBC project by spreading the word to your friends, family, and co-workers. Money raised will help pay for materials and tools needed to drill wells in Isongo, as well as travel and lodging costs for student volunteers.
With your help, EWB-UMBC hopes to raise $20,050 to send a team of 6 students to Kenya to complete this important project.
For more information or to donate, please visit EWB-UMBC's crowdfunding page.
Congratulations to Environmental Engineering graduate student Aditi Bhaskar for receiving a Rising Young Star award at the 2013 Gordon Conference on Catchment Science, which was held Jun 16-21, 2013 in Andover, NH.
Regular conference meetings were preceded by a two-day Gordon Research Seminar, in which graduate students and post-doctoral investigators met as a group to share and discuss their cutting-edge research, build new scientific collaborations and prepare for their interactions with more senior scientists. The goal of the symposium was to promote networking among the young people in advance of the conference in a non-threatening, low-key environment. Students were required to apply in order to be allowed to give oral presentations, and the top three outstanding oral presentations were selected by a vote of all seminar attendees. Rising Young Star recipients were then given the honor of presenting their work to the general conference.
Ms. Bhaskar, who is a research assistant in the Center for Urban Environmental Research and Education (CUERE), did a wonderful job giving her presentation and received congratulations from attendees from around the world, many of whom are senior and well known scientists. CBEE is very proud of her remarkable achievement!