Leveraging Transient Expression

Transient expression is a convenient alternative to developing stable cell lines for bioengineered protein expression

. It is also used for antibody sequencing.These short term protocols involve protein production that can range from just a few days to weeks.

Transient expression helps to achieve high yields in short spans of time and this is especially beneficial in applications like new drug discovery, assay development, high throughput screening and the stage of drug development prior to clinical trials. This is achieved by transfecting the target gene into the producer cells. The protein is then expressed in a few days as the expression plasmid gets lost during cell division. There are several ways to make it more stable.

Transient expression and monoclonal antibody sequencing

While transient expression is used in protein expression, monoclonal antibody sequencing ormAbs is completely different. The mAbs are heterotetramers made up of two heavy and two light chain polypeptides bound by disulfide bonds. To produce full length mAbs using transient expression, one of two things must be done:

Transfect a single plasmid that has both the heavy and light chain coding sequences

Co-transfect individual vectors where each one has either the heavy or light chain coding sequence

If a single plasmid is used, the total DNA concentration in the culture and the ratio of DNA to transfection reagent are critical factors. These factors are influenced by cell type and density and the age of the culture. Of the two methods mentioned above, transient expression from a single plasmid is easier.

At the beginning of the monoclonal antibody sequencing program, the heavy and light chain coding sequences are expressed from plasmids that show all the required genetic elements. Besides determining the optimal total plasmid DNA concentration included in the culture and the plasmid to DNA to transfection reagent ratio, it is important to determine the heavy chain to light chain plasmid ratio to express the full length monoclonal antibody sequence.

Applications for transient expression

A number of applications for these exist in biotechnology and this includes toxicity and affinity studies and activity of the product. Although transient expression is mostly used in drug discovery and preclinical phases, the number of applications is likely to increase as the demand for recombinant proteins in a specific niche increases. Among the three types of cells used, the HEK 293 cells are used in producing recombinant viruses employed in clinical trials. These are also used in biotherapeutics.

Drug development requires, right from the initial stages, a large number of antibodies both in preclinical and clinical studies. The transient expression system chosen depends on various factors such as quantity and quality required and speed at which it can be generated. Transient expression is the preferred method as it generates quick results. Its yields can be managed by optimizing the system to eliminate unwanted indications from the gene and improve it, enabling faster clinical outcomes.

by: Paul Kerr

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