ABSTRACT

Basic Information

Abstract Number: 900 - 7
Author Name: J Christopher Love - Massachusetts Institute of Technology
Session Title: Pittsburgh Analytical Chemistry Award
Event Type: Awards
Event Title: Engineering Single-Cell Bioanalytics for Strain Optimization in Biomanufacturing

Presider Name:Jane Chan
Affiliation:Bechtel Bettis, Inc.

Date: Tuesday, March 19, 2013
Start Time: 10:30 AM (Slot #7)
Location: 114

Abstract Content

Bioprocess development relies on methods to identify optimized cell lines producing heterologous proteins. The establishment of regulatory pathways to enable generic biologic therapeutics—so called biosimilars—may offer an opportunity to increase the accessibility of effective treatments in both the developed and developing world. The cost of goods for manufacturing monoclonal antibodies can be substantially lowered if produced in alternative microbial hosts, but it has been difficult to match the productivity of state-of-the-art mammalian cell lines for monoclonal antibodies.

This talk will describe how an approach for integrated single-cell analysis reveals the diversity and dynamic behaviors in secretory capacity among clonal yeast cells. The methods use arrays of subnanoliter wells to isolate cells (sampled from a reactor) at a density of approximately one cell per well; these arrays loaded with cells then enable high-throughput characterization (10^4–10^5 cells per array) of the secretory capacity among the yeast cells by a technique called microengraving. In particular, the talk will focus on strain characterization and optimization of the methylotropic yeast, [i]Pichia pastoris[/i], which is used to produce biotherapeutics globally, yet lags the productivity of conventional Chinese hamster ovary (CHO) cells for monoclonal antibodies. Single-cell measures of secretion of biologic products from [i]P. pastoris[/i] suggest that there is functional diversity in the productivity among individual cells, and that these populations can shift dynamically. Furthermore, these analytical measures, combined with computational modeling, indicate that transport of folded protein through the secretory pathway is a rate-limiting process in production. Such measures of single-cell distributions and fluctuations offer an alternative approach to optimize the secretory capacity of strains and to monitor changes in process conditions in manufacturing.