The Importance of Temperature Control in Chromatography

The Problem

The operating temperature of a chromatography step used in the purification of monoclonal antibodies (mAbs) was observed to be fluctuating. The various operating temperatures that were observed were having an impact on process performance attributes and product quality resulting in a variable purification step.

The Breakthrough

A temperature assessment of the chromatography step was performed using temperature control apparatus (e.g., a jacketed column and heat exchanger) to assess the impact of temperature on process performance attributes and product quality at a range of operating temperatures.

The Impact

Chromatography steps during the purification of mAbs are employed to capture and purify the mAb while removing and reducing aggregates, residual protein A, host cell protein and host cell DNA. Reliable and consistent performance of these steps is critical to ensure a highly purified product is derived from the process.

Background

Chromatography is temperature dependent and subtle changes of temperature can affect the binding/elution kinetics of proteins. Slower or quicker dissociation time of the mAb from the resin can have an impact on the final eluate volume and protein concentration. This dissociation speed is dependent on the operating temperature of the chromatography step. Temperature consistency can be maintained by using column jackets and heat exchangers to ensure an accurate and precise temperature. To maintain consistency, it is also important to keep all buffers/solutions along with the load material at the set temperature.

Temperature assessment can be overlooked as an operational parameter in chromatography during the process characterization phase. Although variable temperatures may not impact quality attributes, process parameters such as elution volumes or even yield can be affected, leading to loss of product or requiring substantial CAPEX expenditure, such as the purchase of larger holding tanks.

Temperature assessment studies for chromatography unit operations can help define a temperature range at which there is minimal impact to process attributes or product quality and controlling process performance.