Table 1. Analysis of Technology Options for Protein Production

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Table 1. Analysis of Technology Options for Protein Production (acronyms)

Comparative Analyses	Technology Options					Purification
	Cell-Based			Cell-Free	Chemical Synthesis
	E. coli	Alternative Hosts	Homologous Hosts	Cell-Free	Chemical Synthesis
Strengths	Established methods, vectors Renewable Very cost- effective for industrial-scale quantities	Some higher success rates for certain proteins	Codon bias or missing cofactor issues eliminated	Scalable Readily automated Simplified cloning HT screening under readily manipulated conditions Cofactors Labels Production of toxic proteins	Scalable Potential for automation Labels and unusual amino acids incorporated during synthesis	Some tags demonstrated as high throughput, scalable Numerous chromatography reagents available
Weaknesses	Scalability and high-throughput automation	Less developed methods, vectors Cost Not high throughput	Large efforts to develop methods, vectors, strains Scalability and high-throughput automation	Currently only spontaneous disulfide bond formation	Ligations possible at only a small number of amino acid residues Refolding required	Tag removal Tag interference
Development Targets and Needs	More strains, vectors, procedures for difficult proteins	Improved vectors, strains, procedures for difficult proteins	Procedures generalized to engineer uncharacterized microbes	Automation demonstrated Directed disulfide bond formation Difficult proteins	Protein folding problem solved Automated for high throughput	Capability to predict effects of tags Microfluidics Integration with characterization Predictive capability for best purification and storage

June 14-16, 2004, GTL Technology Deep Dive Workshop, Working Group on Genome-Based Reagents

The table above compares and contrasts strengths, weaknesses, and development needs of technologies for use in a high-throughput production environment.