Genetic Engineering and Protein Expression

High level of protein expression is critical for efficient and economical purification of active recombinant proteins. We offer a number of expression systems designed to meet specific goals, ranging from expression of tagged proteins as antigens for antibody generation to production of biopharmaceuticals for clinical use. Our experts can optimize expression by engineering an expression vector with a strong promoter, an efficient ribosome binding site, a high copy number and combining it with an appropriate host. Expression also depends on the gene sequence and on stability, solubility, and folding pathway of the protein product. Accordingly, we will optimize these protein expression parameters when such optimization is necessary.

Genetic Engineering

1. generation of a gene by PCR
2. engineering into an appropriate vector
3. restriction mapping to confirm identity and integrity of a construct
4. DNA sequencing of an insert and consensus sequence derivation
5. mini-plasmid preparation and DNA storage

Expression Systems

1. E. coli and mammalian expression systems
2. commonly used vectors and hosts for expression or systems specified by the customer
3. proprietary high efficiency E. coli expression system offered under separate license for large scale production destined for the clinic

Protein Expression Options

1. stable and transient expression for mammalian expression systems
2. secreted or intracellular proteins for mammalian expression systems
3. cytosolic or periplasmic location for bacterial expression systems
4. tagged or untagged proteins

Expression Assessment

1. soluble vs. inclusion bodies analysis for bacterial expression systems
2. SDS-PAGE, Western, ELISA or functional analysis of cell lysates or conditioned media

Expression Optimization

1. efficient promoter
2. ribosome binding site
3. vector copy number
4. appropriate host
5. growth conditions including media and additives selection, temperature, induction

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