Molecular Biology

Move Your Drug Program Forward Faster
DowpharmaSM practices a rigorous, disciplined approach to process development that advances customers’ drug programs forward faster. For each customer project, we use advanced technologies to aid in the microbial expression of proteins and peptides, and we offer a number of expression technologies, including Pfēnex Expression Technology™, a new Pseudomonas-based system.

Molecular biology services include:
  • Gene cloning
  • Gene design
  • Vector construction
  • Directed strain improvement
  • Gene expression
  • Maintenance of recombinant microbial expression strains

Our scientists work with you to achieve your technical goals.

Cutting-Edge Technologies

In addition to industry-standard techniques, Dowpharma also employs cutting-edge technologies on every customer project to speed strain development and the rapid process development of robust manufacturing programs:

Bioinformatics

  • Full range of computational tools for whole genome analysis and manipulation

Genomics

  • Dow has sequenced and annotated the entire genome of P. fluorescens biovar 1 strain MB214
  • Four other P. fluorescens genomes are publicly available

Genome-Scale Metabolic Modeling

  • Constraint-based, genome-based mathematical model for P. fluorescens

Functional Genomics

  • Transcriptomics
  • Proteomics
  • Metabolomics

A Deeper Look at Functional Genomics: Enabling Rapid, Directed Strain and Process Improvement

Primary Applications of Transcriptomics
Characterizing genes which are involved in:

  • Carbon utilization
  • Protein degradation pathways
  • Secretion pathways
  • Protein folding
  • Disulfide bond formation

Identifying genes which are:

  • Growth-phase regulated
  • Constitutively expressed

Primary applications of proteomics
Analysis of shake flask and 20 liter fermentation experiments:

  • Obtain information about metabolic state of host cells
  • Determine if desired protein or peptide is produced and in what state
  • Protein yield and stability
  • Temporal profile of desired protein production
  • Signal sequence processing

Protein and peptide purification optimization:

  • Identify co-purifying proteins to drive purification processes
  • Steer direct genetic engineering to eliminate interfering proteins

Establishing a Host Strain – An Overview

Dowpharma helps you establish a robust host strain for the production of recombinant proteins and peptides. Below is an overview of the process:

DNA Design and Synthesis

  • Develop cloning strategy
  • Analyze existing gene
  • Synthesize gene; optimize codons if necessary

Strain Construction

  • Design and prepare primers
  • PCR amplify gene and secretion leaders
  • Subclone PCR product containing gene into expression vector
  • Perform DNA sequencing
  • Prepare glycerol stocks

Expression Analysis

  • Perform shake flask fermentations
  • Develop analytical methods
  • Determine yield and quality of protein or peptide
  • Perform purification and analysis
  • Transfer recombinant protein or peptide to customer for evaluation

For more information about Dowpharma’s molecular biology capabilities, contact us.


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