text: Engineering Genes, Vectors, Constructs and Clones - Day 2
  Fourth AnnualEngineering Genes, Vectors, Constructs, and ClonesUpstream Decisions Lead to Downstream SuccessJanuary 21-22, 2013 Day 1| Day 2 | Download Pipeline 4 Brochure TUESDAY, JANUARY 22
  7:15 am Breakfast Presentation: Using Infologs for Multidimensional Gene and Protein Optimization
  Recent advances in gene synthesis and bioinformatics have greatly improved our ability to functionally manipulate gene sequences. Full control over features such as codon bias and mRNA structure allows systematic study of how the gene sequence impacts expression of encoded proteins. Such control also allows efficient exploration of protein sequence variation. We present several studies where informatics-driven gene synthesis is applied for the improvement of heterologous expression of genes in several hosts, including mammalian cell lines, as well as for the engineering of novel protein function.
  Engineering Expression Systems 8:15 Chairperson's RemarksBrian Cain, Ph.D., Key Account Executive, Blue Sky Biotech, Inc. 8:20 Bacterial Chromosome Engineering for Direct Expression of Optimized Proteins
  Joseph Kittle, Ph.D., Assistant Professor, Chemistry and Biochemistry, Ohio University
  This vector- free method features improved gene regulation and optimizes stability and protein yield. Efficient sequence modification techniques targeting host and inserted genes may be used to generate highly selective libraries of bacteria expressing protein variants for analysis and protein production. The process of generating, testing and optimizing expression of variants is simplified and made robust.
  8:50 Construction of Human in vitro Proteome and Its Application
  Naoki Goshima, Ph.D., Senior Research Scientist, Biological Systems Control Team, Biomedicinal Information Research Center, National Institute of Advanced Industrial Science and Technology (AIST)
  We constructed the human ORF clones, Human Proteome Expression Resource (HUPEX), to cover about 80% of entire human genes. HUPEX can be converted to proteins by using wheat germ cell-free translation system (20,000 proteins/week). These proteins were called "in vitro proteome".We introduce the wide range use of HUPEX and "in vitro proteome" in the field of basic and applied research.
  9:20 High-Level Production of Homogeneously N-Glycosylated Heterologous Proteins From a Novel Glyco-Engineered Pichia GlycoSwitch™ Strain by Combined Efforts of Strain and Expression Engineering
  Roland Weis, Ph.D., Head of Operations, VTU Technology
  N-glycosylation of proteins in Pichia pastoris usually consists of branched Man8 – Man14 on the 2 core GlcNAc sugar residues, preventing the application of this powerful host for the production of certain N-glycosylated therapeutic proteins. Using a novel stabilized glyco-engineered platform strain (RCT´s Pichia GlycoSwitch™), resulted in a homogenous N-glycosylation pattern of GlcNAc2Man5 expressed in Pichia pastoris. Several target molecules were investigated for secretory expression levels and homogeneity of N-glycosylation applying VTU Technology´s unique promoter library.
  9:50 Selected Poster Presentation: Development of the epiCHO Transient Expression System for Improved mAb Production
  Trent Munro, Ph.D., Associate Group Leader, Australian Institute for Bioengineering and Nanotechnology, University of Queensland
  10:45 High-Throughput Production at the Macromolecular Therapeutic Development Facility: Multi-Platform Expression Evaluation
  Brandan Hillerich, Ph.D., Head, Protein Production, New York Structural Genomics Consortium, Albert Einstein College of Medicine of Yeshiva University
  The Macromolecular Therapeutics Development Facility at Albert Einstein College of Medicine (MTDF) leverages high-throughput (HTP) proteomic techniques for the development, analysis and production of protein-based therapeutics. During the last 12 months, the MTDF has generated more than 11,000 constructs for more than 5000 protein targets, resulting in over 2500 soluble proteins distributed for functional and structural studies. We highlight our construct design and platform evaluation process.
  11:15 Improved Tools for Expression of Integral Membrane Proteins
  James A. Ernst, Ph.D., Scientist, Departments of Protein Chemistry & Early Discovery Biochemistry, Genentech, Inc.
  Suppression of basal transcription and regulation of transcription activation can avoid cytotoxicity and allow successful expression. Further, proper regulation of translation initiation is vital for folding of many translocated proteins. I will discuss novel transcription control elements with applications to expression of several membrane proteins. Importantly, these promoters allow the successful application of translational regulation to the expression of integral membrane proteins in E. coli.
  » FEATURED SPEAKER 11:45 DNA Aptamers to Detect Subtle Modifications during the Development and Manufacture of Therapeutic Proteins
  Zuben Sauna, Ph.D., Principal Investigator, Laboratory of Hemostasis, Center for Biologics Evaluation and Research, US Food and Drug Administration
  There is a need for detecting conformational changes during the development of a therapeutic protein, following changes in manufacturing processes and for comparing biosimilars and bio-betters to the reference protein. However, there are not many tools to readily and routinely assess conformational integrity. I will discuss in this talk a novel method that uses DNA Aptamers for epitope mapping of therapeutic-proteins and new methods for the development and characterization of a panel of Aptamers for this purpose.