Day 1 :
Keynote Forum
Bevin Page Engelward
Faculty at the Massachusetts Institute of Technology
Keynote: Novel Cytotoxicity and Broad-Spectrum Genotoxicity Platforms
Biography:
Bevin Page Engelward graduated from Yale University in 1988 and from the Harvard School of Public Health in 1996. She continued at Harvard for a one year postdoc, after which she joined the faculty at the Massachusetts Institute of Technology. She is currently Professor of Biological Engineering, Deputy Director of the Center for Environmental Health Sciences, and Director of the MIT Superfund Research Program Center. The main interests of Prof. Engelward are DNA damage and repair, and development of novel technologies relevant to cancer etiology and drug development. In particular, she leads efforts to exploit photolithography to create cell microarrays that serve as a platform for novel toxicity and genotoxicity assays. Most recently, a high throughput cytotoxicity assay has been developed wherein toxicity is measured by a change in the distribution of microcolony sizes. She has also helped to develop a higher throughput DNA damage assay that is based on the comet assay. The “CometChip” has been further developed to detect diverse classes of DNA damaging agents, including potentially carcinogenic bulky lesions. In addition to studies of cytotoxicity and genotoxicity, her laboratory was the first to develop a mouse model wherein mutations (caused by homologous recombination) can be detected in situ via fluorescence. This model has been used to reveal genetic and environmental factors that modulate susceptibility to large scale sequence rearrangements.
Abstract:
Toxicity and Genotoxicity testing are fundamental to drug safety and drug development. Here, we leverage cell microarray technology to create a robust and highly sensitive cytotoxicity platform and a broad-spectrum genotoxicity platform.
• Quantification of cell viability is one of the most fundamental and broadly used endpoints in the life sciences. The gold standard is the colony forming assay. While the assay has an impressive dynamic range (over several orders of magnitude), it is relatively low-throughput (10-21 days), laborious and requires large dishes/high volumes of media, thus requiring large amounts of test compounds.
• Microtiter cytotoxicity assay have thus been developed, including the XTT and the CellTiter-Glo assays. The XTT assay suffers from low sensitivity, and the CellTiter-Glo assay is subject to artifacts due to its indirect measure of cell viability.
• To overcome these limitations, we developed the MicroColonyChip (uCC) assay, which directly measures the ability of cells to divide (like the gold-standard colony forming assay), but with the scale and speed of microtiter assays. Microcolonies grow in a microarray and toxicity is derived using a novel metric, namely the change in the distribution of microcolony sizes. The result is an exquisitely sensitive assay that is robust to artifacts.
• For genotoxicity testing, the comet assay is a commonly used approach. We recently developed a higher throughput version of the comet assay that exploits a cell microarray. The “CometChip” is more than 1000X faster, far more sensitive and includes automated data analysis. Further, we broadened the spectrum of detectable lesions to include bulky lesions, a class of DNA damaging agents that has the potential to cause cancer.
Accurate cytotoxicity and genotoxicity testing hold a central role in drug development. Having reliable and sensitive assays enables identification of untoward deleterious effects of drug candidates, providing immense savings by narrowing the candidate pool. Further, cytotoxicity and genotoxicity assays are also pivotal for development of novel DNA damaging chemotherapeutics, the mainstay of cancer treatment today.
- Industrial Biotechnology
Session Introduction
Samuel Nii Odoi Yemofio
University of Cape Coast, Ghana
Title: Investigation of itaconate metabolism in Cupriavidus necator H16
Biography:
My strong interest for Biotechnology developed during my undergraduate program to extract DNA of a plant species in the University of Cape Coast, Ghana. I decided to further my studies at the University of Nottingham-UK, with my research at the Centre for Biomolecular Sciences to improve my laboratory skills and experience and contribute to solving issues regarding climate change the future of the environment. My experiences as a teaching assistant and during my research project have enabled me to develop a strong passion to enhance my career in microbial biotechnology. My professional experiences have revealed the complexity of managing and maximizing my time and interpersonal skills to achieve my set targets. My expertise in evaluation and passion is in microbial biotechnology to identify new approaches and improve chemical, fuel and energy production. His open and contextual evaluation model based on responsive constructivists creates new pathways for improving the energy sector.
Abstract:
Statement of Problem: Recent challenges of pollution and climate change in our environment stems from the over dependency on fossil fuel through the extraction, processing and exploitation for petrochemicalbased products. This has caused severe havoc to the environment and its natural habitats, leading to deaths and displacements into unfavourable conditions. Researchers in the US Department of Energy (DoE) in 2004 identified itaconate, one of the twelve attractive platform chemicals, as a potential chemical suitable for bio-based industrial products using biological routes. Previous research has also shown that itaconate has the potential to replace petroleum-based products such as petrochemicalbased acrylic and methacrylic acid; and detergents, surface active agents and biosynthesized plastics for industrial applications with bio-based products. This can be achieved through biological or chemical conversions and be subsequently converted into several high-value biobased chemicals and materials from biomass. Research also discovered that itaconate is naturally produced by microorganisms such as Candida sp., Ustilago madis and Aspergillus terreus although many microorganisms have been genetically engineered for the biosynthesis of itaconate. It is therefore necessary for current generation to identify various sustainable and cleaner processes for chemical, fuel and energy production. HPLC was used to estimate the concentration of itaconate consumed. The purpose of this research was to identify the genes involved in itaconate metabolism and abolish its metabolism. Methodology & Theoretical Orientation: To investigate itaconate metabolism on host organism Cupriavidus necator H16, growth of mutants was observed using itaconate as sole carbon source. Findings: Single, double and triple knock-outs of ict genes involved in itaconate conversion to itaconyl-CoA (itaconate-CoA transferase activity) were generated. Growth and itaconate consumption assays were performed establishing that only H16_RS22140 gene is clearly involved in itaconate metabolism. Recommendation: This study revealed that other genes can be involved in itaconate degradation and therefore further research to investigate function of these genes is required.
Dr. Sourav Majumdar
Deputy Manager in Serum Institute of India Pvt. Ltd, Pune
Title: Assessment of N-linked Glycosylation and physico-chemical properties on biological activity of anti-HER2 molecule
Biography:
Dr. Sourav Majumdar serves as a Deputy Manager in Serum Institute of India Pvt. Ltd, Pune. He has developed his core expertise in the area of Bioanalytical method development in proteomics platform. He is leading the group of biosimilar especially dealing with recombinant monoclonal antibody. After completion of his Ph.D. degree he joined EPR Centre for cancer research, Hyderabad as ADL lead. He developed several analytical methods for routine and extensive characterization of mAb molecule. The documents were submitted for Regulatory authority such as RCGM, DCGI for product registration. Formerly, he worked in Intas Bio Pharmaceutical R&D stability division as Senior Research Associate. He then joined for USV and was assigned to work on various recombinants Biosimilar Therapeutics with several analytical challenges especially with anticancer molecule. He joined as a Junior Research fellow in MBBT department for Ph.D. His research was on a “Novel Fibrinolytic Enzyme and a comparison with commercially available Cardiovascular Drugs under in vitro and in vivo conditions”. He has his majors in Bio-Chemistry from University of Pune, 2004. He is a recipient of “National Level fellowship” and “Ratan Tata fellowship”. He was awarded the best speaker from Garware Chemical Association. He attended several National and International Conferences as a speaker. He is a life member of Indian Science Congress and Society for Biological Chemist. He published a series of Research Article in reputed International Journals. In addition, he is also a playing vital role for establishing a GMP facility.
Abstract:
Background: Glycan moieties and charge variants decisively influence the efficacy, potency of the anti-HER2 antibody. A minor change in glycan profile of anti-HER2 molecule during post translational modification showed an impact on its biological activity, Pharmacokinetics (PK) property of the molecule. The complexities in glycan pattern of anti-HER2 molecule further encourage the need of new analytical challenges in evaluating the comparability with innovator molecule in recent scenario. Aim / Objective: A comparison of glycan profiling and binding kinetics including charge variant heterogeneity analysis of anti HER2 molecule with innovator molecule was aimed for the current investigation. Methodology: In the current investigation, The PNGase F digestion of anti HER2 molecule followed by UPLC analysis under Normal phase liquid chromatographic conditions using a platform approach to establish the chromatographic profile of various Glycan residues such as G0, G0F, GIFa and GIFb, G2F. Further, binding with HER2 receptor under in vitro conditions through Biocore T200 analysis was evaluated to demonstrate the binding kinetics. Additionally, charge variant heterogeneity was subjected as a part of extensive characterization to evaluate the anti HER2 molecule in comparison with innovator molecule. Results and Discussion: Anti HER2 molecule with a mass of 148 kDa was purified and a comparison against innovator molecule in glycan profiling was identified by predominant residues, which represent 90% of the glycosylation profile of the Anti HER 2 molecule. The terminal galactosylation and afucosylation of glycan residues influences the CDC and ADCC activity, respectively. The variability in glycan profile can thus affect the biological activity. Further, the high mannose content in anti Her2 molecule showed the early clearance and provides strong impact on its PK. Additionally, binding with HER2 receptor under in vitro conditions through Biocore T200 analysis was evaluated to demonstrate the binding kinetics. The dyanamic binding nature of anti HER2 molecule (6.21X10-10 M) and innovator molecule (6.95 X10-10 M) showed the closeness of KD value. Moreover, the weak binding of acidic variants with HER2 receptor also provides an impact on ADCC activity and hence, charge variant analysis was shown to be equally important to demonstrate the efficacy of the anti Her2 molecule. Acidic variant (K0, deamidated Asn) and basic variants (K1, K2) were determined by cation exchange chromatography (CEX) and percentage of acidic variant (<35%) were evaluated based on separation on cation exchange column in UHPLC system. The comparisons was carried out based on response of reference standard peaks under identical condition and determine the percentage of acidic variants. Conclusion: Overall, the efficacy and comparability aspects of the in-house anti-HER2 molecule were evaluated against the innovator’s molecule and confirm its biosimilarity
Dr.Naila Rozi
Associate Professor Computer science Department at Sindh Madrassatul Islam University
Title: Analysis of Risk in CVD VIA Human Genetics & biomedical equipment
Biography:
Dr.Naila Rozi is working as Associate Professor Computer science Department at Sindh Madrassatul Islam University. She is working in Association with Dean Prof. Dr. Syed Asif Ali. She has twenty years teaching experience in public sector as well as in private sector with Dean Prof .Dr .Javed .H.Rizvi Sir Syed University of Engineering and Technology. Dr.Naila hold PhD in Applied Mathematics subject of operation Research, dissertation on “A Mathematical Approach to Geometry of DNA ”under supervision of Pro Vice Chancellor University of Karachi, Prof .Dr Nasir Uddin Khan. She is HEC Registered Supervisor in Computer Science. She is in Committee Member of CENSUS-2017 PK, She is a senior member of WSSET,NSP,NEHA,ISEIS Canada, AAAI ,ISOSS,ASNR Pakistan ,IACSIT,IASED &APMS. She is a member of editorial board in “Pakistan Journal of Statistics” and Neurographic Journal. She has more than twelve research publications in journals and conference proceeding. She had been training workshop of SPSS as a trainer in SMI, University & SSUET. She is an author of IEEE, She attended many International and National conferences to presented research papers in Dubai, American University Cairo &Qatar University Doha and China.
Abstract:
Statement of the Problem: In applied Biomedical Engineering discipline of human factors analysis is a complex and evolving study in cardiac surgery. Some realistic effort to reduce human error arose with the observational nature of human factors engineering we can take to analyze Risk Theory in cardiac surgery .According report from ECRI, here are the 10 riskiest areas which we have to analyze .Infusion errors which may be deadly to Patient. One big issue that can slip through the cracks is “IV free flow”. Secondly inadequate cleaning of complex reusable instruments which recent rash infections attributed to the reuse instruments after sanitizing and staff should be regularly reminded of the correct cleaning protocol. Third missed ventilator alarming Robotic Surgery may also be associated with higher costs and additional risks.
Risk Analysis in CVD can be monitor through Bayesian Analysis to integrate independent dataset, Bayes factor as a function of SNP in the CHD population. To test robustness of Bayesian analysis, we examine two tests of the sensitivity ,namely to low significance data sets. we exclude the data sets with comparatively small sample sizes, we also exclude large Bayes Factor
Methodology:
We use a Bayesian spatial model to estimate CVD mortality by ward ,sex and age group for period of 2012-2018 in Pakistan.The number of deaths in each ward –period-sex-age group was specified using a Poisson model. The Poisson model estimates mortality in each ward.
- Industrial & Microbial Biotechnology
- Pharmaceutical Biotechnology
Session Introduction
Dr. Nawab Ali
faculty member in the Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Pakistan.
Title: Molecular Characterization and Growth Optimization of Protease Producing Pseudomonas aeruginosa strain (IBC-2)
Biography:
My research and teaching interests have been fostered by 12 years of undergraduate and postgraduate teaching and mentoring as faculty member in the Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Pakistan. My PhD and current research interests are focused upon protein purifications and extremophilic enzymes. I had one project worth 0.5 million funded by Directorate of Science and Technology, Peshawar entitled as “Large scale purification of industrially important proteases utilizing the charge based and hydrophobic interaction based chromatography”. This project is completed and data is published in well reputed journals. The second project has been funded by NRPU-HEC to explore extremophilic microbes collected from desert, hot springs and salt mines in order to produce enzymes which can be used in harsh laboratory conditions. These enzymes will be feasible in extreme laboratory conditions such as temperature, pH and high salt concentrations. This may result to identify novel bacteria/fungi with high expression quality and may result in patents
Abstract:
Microbial proteolytic enzymes are used to hydrolyze proteins in various bio-industrial applications but most of the enzymes are not active in extreme conditions of temperature, pH and salt concentrations. This increases the demand to explore enzymes to show activity at such extreme environments. Exciting findings have been described about the extremophilic bacteria isolated first time from coal mines of Orakzai Agency, Pakistan, having protease producing properties. Among various extracted isolates, â“6 isolates were purified based on showing proteolytic activity using 2% skim milk (SM) agar media at 37oC for 24 h, to identify various morphological and biochemical characteristics. The isolate, IBC-2 showed the maximum zone of activity (21mm) and was identified as Pseudomonas aeruginosa (JCM 5962) using 16S rRNA gene sequencing. The strain IBC-2 exhibited significantly (P ≤ 0.05) maximum growth and protease production (335 U/ml) at 37°C and at pH 7 after 24 h of incubation. The strain was also significantly (P ≤ 0.05) more active for protease production at neutral pH (435 U/ml) than at high or low pH which reflected the neutral nature of the bacteria. In conclusion, the strain IBC-2 has the highest protease producing ability with distinctiveness, which can
- Applied Biotechnology