2^nd International Conference on Genetic and Protein Engineering November 14-16, 2016 Atlanta, Georgia, USA

Nicholas Agyepong holds MSc Biotechnology and Phil Pharmaceutical Microbiology degrees from Kwame Nkrumah University of Science and Technology in Ghana. He was a Research Assistant between 2005 and 2007 in the Parasitology laboratory, Noguchi Medical Research Institute, Accra. He later became Lecturer and went through the ranks to become a Head of the Department of Dispensing Technology at Sunyani Polytechnic in Ghana. He is currently PhD Pharmaceutics student at the University of Kwa-Zulu Natal in South Africa, under the supervision of Prof. Sabiha Y Essacks.

Antimicrobial resistance is currently a major scientifi c concern both in hospital and community settings globally due to the increasing rate of numbers of bacterial strains acquiring resistance to many relevant antibiotics Th e bacterial pathogens that are of great medical importance and associated with outbreaks of highly drug resistant strains include the ‘ESKAPE’ pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa andEnterobacter spp.). Th e resistance mechanisms include mutations in penicillin binding proteins, effl ux mechanisms, alterations in outer membrane proteins and the production of hydrolyzing enzymes. However, the most common resistance mechanismto β-lactams antibiotics is the production of β-lactamases among its Gram-negative isolates. Some important β-lactamases contributing to therapeutic failure include extended-spectrum β-lactamases, metallo-β-lactamase, AmpC β-lactamase and carbapenemases such as Klebsiella pneumoniae carbapenemases. With the global surge in the occurrence of these resistantepisodes, prompt detection is necessary for implementation of strict infection control policies and treatment with alternative antimicrobials. In spite of this, relatively little governmental, funding and academic attention has been received in Ghana and the West Africa sub-region. Th e paucity of data on the prevalence of resistant among these clinical species is an index of limited research carried out so far; hence, the study to determine the resistance factors and mechanisms among the abovementioned clinical isolates. Th us providing quality data on resistance prevalence become a baseline for further research and comparative studies. Phenotypic detection of β-lactamases and confi rmation was done using CLSI, (2014) guidelines. Detection of β-lactamases multiple genes was done using whole sequencing technology.

Tanya Hundal is a fi nal year graduate student at Department of Chemistry, University of South Carolina and is working with Dr. John J Lavigne.

Cancers of colon and prostate, though treatable, require early detection to improve patient outcomes. Current diagnosis methods e.g. visual inspection and biopsies are somewhat subjective, thus decreasing accuracy. Alternatively, blood-based tests measuring specifi c biomarkers (like CEA and PSA) are associated with high false-positive rates and are more useful for monitoring post-treatment patient health, thus driving eff orts to identify better screening and diagnostic techniques. Abnormal glycosylation of integral membrane and secreted glycoproteins is known to take place at the onset of many diseases, including cancer, and presents as the over, under or new occurrence of certain glycans. Th e aim of this study is to design synthetic lectins (SLs) that recognize cancer associated glycans (CAGs). Each CAG produces a unique response pattern with an array of crossreactive SLs. Further, the ability of an array of SLs to discriminate cell lines based upon their diff erence in metastatic potential, demonstrates an alternative approach to detect colon and prostate cancers by looking at global changes in glycosylation. The utility of these SLs was demonstrated using purifi ed glycoproteins, since these glycoproteins expressed similar glycans which are also present in CAGs. Previously, using statistics, an array of 5 SLs could distinguish between 4 human colon cell lines based on their metastatic potential with 89% accuracy. Further SL design modifi cations have been carried out and correlations between structural modifi cations and activity have been established. Based on these preliminary examinations, several new SLs have been identifi ed targeting human colon and prostate cell lines. An extended array incorporating these new SLs, can discriminate between normal, cancerous non-metastatic and cancerous metastatic secreted proteins of diff erent tissue types even better. Ongoing work is focused on investigating the binding interaction between SLs and glycans/proteins to enhance our understanding of the system in order to improve upon current SL structures.

In 2013 he obtained master degree in biotechnology at the Jagiellonia University in Krakow, Poland. The master project was focused on developing a novel scFv expression system. He gained new experience in Fab synthetic library generation and phage display technology during one-year internship program at The University of Chicago, USA. In October 2014 he became a PhD student of Structural Biology Brussels, Belgium. From that point he has had the opportunity to participate in a number of research topics as a Jan Steyaert’s laboratory member. Most of them are focused on protein engineering and different methods of protein display of yeast, bacterial or phage surface. Recently he has been focusing on developing a new yeast display technology for Nanobody discovery platform.

For decades, antibodies have played a central role as research tools and more recently also as therapeutic agents. Their impact on research and medicine is still growing. Nanobodies (Nbs) are the small (15 kDa) and stable single-domain antibody fragments harboring the full antigen-binding capacity of the original heavy chain-only antibodies that naturally occur in camelids. Over the last years, the Steyaert lab pioneered the use of Nbs in structural biology. Nbs are encoded by single gene fragments, are easily produced in microorganisms and exhibit a superior stability compared to the derivatives of conventional antibodies like Fabs or scFvs. Because of their compact prolate shape, Nbs expose a convex paratope and have access to cavities or cleft s on the surface of proteins oft en inaccessible to conventional antibodies. We have demonstrated that Nbs are exquisite antibodies that can bind the most diffi cult targets including membrane proteins, transient multiprotein assemblies, transient conformational states and intrinsically disordered proteins. Our Nanobody discovery platform utilizes routinely phage display approach and more recently, shift ing to the yeast display technology. In yeast display system each yeast cell is capable of displaying up to 100,000 copies of one Nb clone. Th ereaft er each yeast cell can be stained and analyzed for binding of the fl uorescently labeled antigen. Fluorescence-Activated Cell Sorting (FACS) allows the fi ne-resolution separationof cells by measuring a number of distinct optical parameters in parallel. We are presenting our modifi ed yeast display system, where displayed Nbs are fl uorescently labeled in the one-step orthogonally staining. Features of selected Nbs can be fi rst easily characterized using the power of FACS, without a need of recloning, expression and purifi cation of each Nb. Th e Nanobody discovery platform is opened for diverse collaboration partners through Instruct Integrating Biology.

Frederic Cadet is Vice President in Research & Development of the company Peaccel. He has a PhD in Protein Engineering, Data mining and Biosimulation. From 2004 to 2008, as an Executive School, University & Research Commissioner, he managed a budget of 1.3 billion Euros and was responsible for 32,000 employees. He is Former Chairman of the ERA Nets (European Research Area Networks) NetBIOME. He has developed pioneering research activities in bioinformatics. He is author of over 70 publications and Referee for 17 international scientifi c journals. He is Organizing Committee Member for 2nd International Conference on Genetic and Protein Engineering, November 14-16, 2016 Atlanta, USA.

We present a strategy that combines wet-lab experimentation and computational protein design for engineering polypeptide chains. Th e protein sequences were numerically coded and then processed using Fourier Transform (FT). Fourier coeffi cients were used to calculate the energy spectra called "protein spectrum". We use the protein spectrum to model the biological activity/fi tness of protein from sequence data. We assume that the protein fi tness (catalytic effi cacy, thermostabilty, binding affi nity, aggregation, stability) is not purely local, but globally distributed over the linear sequence of the protein. Our patented method does not require any protein 3D structure information and fi nd patterns that correlate with changes in protein activity (or fi tness) upon amino acids residue substitutions. A minimal wet lab data sampled from mutation libraries (single or multiple points mutations) were used as learning data sets in heuristic approaches that were applied to build predictive models. We show the performance of the approach on designed libraries for 3 examples: Enantioselectivity,thermostability and binding affi nity. We can screen up to 1 billion protein variants (10^9).

Frederic Cadet is Vice President in Research & Development of the company Peaccel. He has a PhD in Protein Engineering, Data Mining and Biosimulation. From 2004 to 2008, as an Executive School, University & Research Commissioner, he managed a budget of 1.3 billion Euros and was responsible for 32,000 employees. He is Former Chairman of the ERA Nets (European Research Area Networks) NetBIOME. He has developed pioneering research activities in bioinformatics. He is author of over 70 publications and Referee for 17 international scientifi c journals. He is Organizing Committee Member for 2nd International Conference on Genetic and Protein Engineering, November 14-16, 2016 Atlanta, USA.

The knowledge of organisms and their metabolic pathways allowed constructing biological systems for the production of chemicals and pharmaceuticals such as antibiotics and biofuels. Synthetic biology expands the number of these biological systems by the assembly of artifi cial metabolic pathways, called synthetic pathways, not present in natural organisms. Synthetic pathways could be integrated in modifi ed micro-organisms or in biocatalyst systems. A biocatalyst system is an in vitro assembly composed only of purifi ed enzymes and metabolites that are useful for the production of a desired metabolic compound through a biochemical reaction network. Th is in vitro assembly, as compared to cellular system, has several advantages, such as the production of only desired metabolites and a great engineering fl exibility. We explored an in silico approach to identify and analyze new biocatalyst systems for the production of target metabolic compounds. Th is approach proceeds in several steps. The first step is the enumeration of several biocatalyst systems that could synthesize a target product from a desired starting substrate. Next, a selection based on several criteria is applied to choose a biocatalyst system among the group of biocatalyst systems identifi ed in the enumeration step. The last step is the modeling of the selected biocatalyst system to evaluate the production rate and the yield of the target product. This communication explains in more detail the modus operandi for the different steps of our in silico approach.

Lori A Somner is a fi nal year PhD candidate at the University of Sheffi eld, Chemistry Department, based in the UK (2014-2018). This is also where she was also awarded her Master’s degree in Chemistry (MChem, 2010-2014). She has expertise in chemistry and chemical biology, specifi cally nanotechnology and biomedical applications.

Magnetotactic bacteria are a model organism for magnetite biomineralization and represent a growing research area due to the wide range of applications for the magnetic nanoparticles they can produce. An organelle unique to these bacteria, called the magnetosome, produces a single crystal of magnetite enveloped inside a lipid membrane. A host of specialized proteins are involved in the production and regulation of the magnetosomes, they are capable of synthesizing high quality, uniform, magnetic nanoparticles under ambient conditions. Several key proteins have been identifi ed from these bacteria which display activity in synthetic magnetite precipitation reactions where they improve both the homogeneity and morphology of the crystals. However, these proteins are transmembrane proteins which mean they are diffi cult to produce and characterize.Our research aims to unlock the mechanisms, at the molecular level, through which these proteins control biomineralization.We do this by studying the hydrophilic loops linking their transmembrane helices using both free peptides and constrained stem loop coiled coil scaff olds. Th is approach generates novel biomimetic reagents for precision nanoparticle synthesis, and overcomes the solubility issues of the full membrane protein. Recently, the use of artifi cial protein scaff olds has become an area of intense interest. Th e use of these small, robust, monomeric proteins has become a viable alternative to the much more complex and expensive antibodies. In addition to the coiled coil scaff old, we also utilized a magnetite interacting Adhiron (MIA) scaff old. In the presence of an active loop sequence they are capable of controlling the nucleation and growth phases of magnetic nanoparticle crystallization. Th e structural limit to conformational changes ensures binding is both sequence and structurally dependent, superior to an unstructured peptide. Th e scaff olds may also be used for the attachment of dyes, antibodies and drug molecules to magnetic nanoparticles-making these assemblies possible alternatives to antibody-drug conjugate systems.