Cambridge Healthtech Institute’s 12th Annual

Engineering Genes, Vectors, Constructs, and Clones
( 基因、载体、建构物、植株的开发 )

Exploring Tools and Developing Strategies to Expand the Protein Expression Toolbox


Part of the Biotherapeutic Expression & Production pipeline

The demand for high-quality biotherapeutic proteins has never been greater. Many variables still must be considered during the cell-line development process, including verification and sequence analysis of the gene or protein of interest, codon optimization, vector construction, and clone/host selection – a time-consuming and expensive process. Additionally, protein expression scientists are now exploring new engineering tools, such as synthetic biology and systems engineering. Ultimately, these tools must be weighed against traditional expression and production strategies to achieve the desired quantity and quality. Cambridge Healthtech Institute’s 12th Annual Engineering Genes, Vectors, Constructs, and Clones conference continues the tradition of applying effective engineering strategies for protein expression and production research leading to functional biotherapeutic products. Learn from seasoned, savvy researchers as they share their real-world experiences, applications, and results.

Final Agenda


4:00 - 6:00 pm Pre-Conference Registration


7:00 am Registration and Morning Coffee

Exploring Expression Tools

9:00 Organizer’s Welcome Remarks

Mary Ann Brown, Executive Director, Conferences & Team Lead, PepTalk, Cambridge Healthtech Institute

9:05 Chairperson’s Opening Remarks

Harun Rashid, PhD, Senior Principal Scientist, Molecular Technology, Ambrx, Inc.



9:10 High-Throughput Expression of Functional Proteins in a Microarray Format

Josh LaBaer, MD, PhD, Executive Director, Biodesign Institute, Arizona State University

The Nucleic Acid Programmable Protein Array uses printed cDNAs as templates to produce full-length proteins in situ, enabling high-throughput biochemical testing of thousands of well-folded proteins simultaneously. The recombination-based vector system allows users to routinely execute the automated transfer of thousands of genes into useful protein expression vectors overnight. These methods are now extended to include decorating the proteins with post-translational modifications, including glycosylation, acetylation, citrullination and others.


9:50 Codon and Codon-Pair Usage Tables (CoCoPUTs): Facilitating Genetic Variation Analyses and Recombinant Gene Design

Chava Kimchi-Sarfaty, PhD, Deputy Associate Director for Research, Office of Tissues and Advanced Therapies, CBER, FDA

We have created for codon and codon-pair usage tables (CoCoPUTs), genome and tissue specific, a new and regularly updated website that encompasses the previously generated High-Performance Integrated Virtual Environment codon usage tables (HIVE-CUTs) that has also been expanded to include codon-pair usage and dinucleotide statistics. The use of this tool will be discussed specifically for therapeutics’ design.

10:20 Networking Coffee Break

10:45 Cell-Free Protein Synthesis from Genomically Recoded Bacteria Enables Multisite Incorporation of Non-canonical Amino Acids

Antje Krüger, PhD, Postdoctoral Fellow, Michael Jewett Laboratory, Department of Chemical and Biological Engineering, Northwestern University

Cell-free protein synthesis has emerged as a powerful approach for expanding the range of genetically encoded chemistry into proteins. We recently established a bacterial cell-free protein synthesis platform based on genomically recoded Escherichia coli lacking release factor 1 that enables both high-yield protein synthesis and incorporation of multiple, identical non-canonical amino acids. In this talk, we will discuss the development of this platform and its use in engineering the translation machinery.

11:15 Expression Optimization of an Antibody Fab Fragment in Escherichia coli with Non-Native Amino Acid (NNAA) Incorporated

Harun Rashid, PhD, Senior Principal Scientist, Molecular Technology, Ambrx, Inc.

In this study, expression of a “difficult-to-express” antibody Fab fragment with a non-native amino acid (NNAA) inserted was systematically optimized by expression vector engineering. After the various genetic elements on expression vector were tested individually, the beneficial ones were then combined into a single expression vector, which resulted in significant improvement of Fab titer over the starting strain.

11:45 Optimizing Gene Sequences for Improved Protein Expression in Industrial Microorganisms

Tomoshi Kameda, PhD, Senior Research Scientist, National Institute of Advanced Industrial Science and Technology

12:15 pm Sponsored Presentation (Opportunity Available)

12:45 Session Break

12:55 Luncheon Presentation I to be Announced

1:25 Luncheon Presentation II (Sponsorship Opportunity Available)

Gene Editing

2:00 Chairperson’s Remarks

Jeffrey Barrick, PhD, Associate Professor, Molecular Biosciences, The University of Texas at Austin

2:05 Transposon-Encoded CRISPR–Cas Systems Direct RNA-Guided DNA Integration

Samuel H. Sternberg, PhD, Assistant Professor, Department of Biochemistry and Molecular Biophysics, Columbia University

Conventional CRISPR–Cas systems maintain genomic integrity by leveraging guide RNAs for the nuclease-dependent degradation of mobile genetic elements. We uncovered a remarkable inversion of this paradigm, in which bacterial transposons co-opt nuclease-deficient CRISPR–Cas systems to catalyze RNA-guided integration of mobile genetic elements into the genome. This discovery of a fully programmable, RNA-guided integrase lays the foundation for genomic manipulations that obviate the requirements for double-strand breaks and homology-directed repair.

2:35 Defending Engineered Bacteria against Evolutionary Failure

Jeffrey Barrick, PhD, Associate Professor, Molecular Biosciences, The University of Texas at Austin

Mutations in engineered DNA sequences can occur rapidly and infiltrate processes with ‘broken’ cells. Selfish genetic parasites–such as transposons and prophages–are a major source of inactivating mutations in bacterial genomes. In this talk, I will describe the broad-host-range CRISPRi-ME system that can be added to a bacterial cell to repress these mobile elements and other strategies for creating chassis cells with lower-than-natural mutation rates to stabilize their functions.

3:05 Find Your Table and Meet Your BuzZ Session Moderator

3:15 BuzZ Sessions with Refreshments

Join your peers and colleagues for interactive roundtable discussions.

Click here for more details

Vector Design

4:30 A Nuclear Genetic Sensor to Measure and Optimize Delivery of Non-Viral DNA into Human Cells

Karmella Haynes, PhD, Assistant Professor, Wallace H. Coulter Department of Biomedical Engineering, Emory University

The small fraction of DNA that reaches the nucleus during non-viral gene delivery is often silenced by mechanisms that are not well understood. Viral transduction is a robust alternative, but has critical limitations, such as cargo size, and side effects, such as immunogenicity. We developed a genetically encoded sensor to track the fate of unlabeled non-viral DNA in live cells and to support efficient screening for interventions to improve gene delivery.

5:00 Development of Pediatric Gene Therapy Using Nuclease-Free Genomic Editing Technology, GeneRide

Jing Liao, PhD, Associate Director, Discovery Biology, LogicBio Therapeutics

GeneRide is a promoterless, nuclease-free genome editing technology. Combined with highly liver-tropic AAV vectors, GeneRide harnesses the natural process of homologous recombination to integrate the therapeutic gene site specifically into the Albumin (Alb) locus in a non-disruptive manner. Following GeneRide treatment, expression of the therapeutic gene is linked to that of Albumin via a 2A peptide and can be applied to treat the pediatric disease.

5:30 Bioresponsive Liposomes – Modulating the Trigger to Improve Site-Specific Delivery

Francis C. Szoka, PhD, Professor of Bioengineering, Therapeutic Sciences and Pharmaceutical Chemistry, University of California, San Francisco

Four decades since the original publications of pH-triggered liposomes, improved chemistries and a much better understanding of the cell biology of endocytosis/phagocytosis have provided components that can be used for the intracellular delivery of macromolecules. I’ll describe modified lipids that can be incorporated into liposomes and that are activated by changes in pH, redox potential, or phosphatase activity during the delivery phase to enable intracellular content delivery.

6:00 - 7:15 Welcome Reception in the Exhibit Hall with Poster Viewing

7:15 Close of Day


8:15 am Registration and Morning Coffee

CHO Cell Line Engineering

8:45 Chairperson’s Remarks

Nathan E. Lewis, PhD, Associate Professor, Department of Pediatrics, University of California, San Diego

8:50 Predicting Growth Phenotypes of Mammalian Cell Culture Using Genome-Scale Metabolic Models

Nathan E. Lewis, PhD, Associate Professor, Department of Pediatrics, University of California, San Diego

9:20 Model-Driven Process Development for Enhanced Bioprocessing

Meiyappan Lakshmanan, PhD, Research Scientist & Group Leader, Systems Biology, Bioprocessing Technology Institute, A*STAR

Chinese hamster ovary (CHO) cells are the preferred choice for biotherapeutic protein production. However, ensuring consistent high product quality remains a major challenge. The availability of the CHO genome sequence has enabled the development of genome-scale models (GEMs) to examine the metabolic signatures of CHO cells upon varying bioprocess conditions. This talk will show how the genome-scale models can help process development by characterizing key bottlenecks in media formulations and propose targets for media/feed optimization.

9:50 Coffee Break in the Exhibit Hall with Poster Viewing

Cell-Free Systems

11:00 Democratizing Cell-Free Protein Synthesis: Improving Access for Broad Bioengineering Applications

Javin P. Oza, PhD, Assistant Professor, Chemistry & Biochemistry, California Polytechnic State University, SLO

Cell-free protein synthesis (CFPS) is a platform technology that leverages cell extracts for the on-demand production of proteins. CFPS supports discovery by obviating the barriers presented by the cell, enabling scientists and engineers to rapidly characterize genes, constructs, and clones associated with functional genomics, ‘difficult’ proteins, and metabolic engineering. I will discuss our efforts to reduce the barriers to implementing CFPS to enable broad applications in bioengineering.

11:30 Using Human Blood Extracts as a Renewable Resource to Produce Recombinant Proteins

David Burgenson, PhD Candidate, Center for Advanced Sensor Technology (CAST), University of Maryland Baltimore County

One of the early in vitro translation cell-free protein expression systems used rabbit reticulocytes (immature red blood cells) as a source of cells to produce active cell-free extract. Using this system as a source of inspiration, our group began looking for cells present in human blood that could be used to produce translationally active cell-free extract.

12:00 pm Presentation to be Announced

12:30 Session Break

12:40 Luncheon Presentation to be Announced

1:10 Close of Engineering Genes, Vectors, Constructs, and Clones Conference


5:45 - 8:45 Recommended Dinner Short Course*

SC6: Assembling an Effective Toolbox of Expression Systems to Support Your Drug Discovery Efforts

Richard Altman, Field Application Scientist, Protein Expression, Biosciences Division, Life Sciences Solutions Group, Thermo Fisher Scientific
Henry C. Chiou, PhD, Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific
Dominic Esposito, PhD, Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research


*Separate registration required

* 活动内容有可能不事先告知作更动及调整。

Choose your language