08 Downstream Processing and Biomanufacturing Free Online FlashCards

What is the typical downstream-processing sequence?

The typical sequence is recovery and clarification → capture → purification → polishing → formulation.

How does product location affect recovery?

A secreted product is recovered from the culture liquid. An intracellular product requires cell disruption before recovery.

Which operations commonly clarify culture broth?

Centrifugation and depth or membrane filtration clarify culture broth by separating cells and other bulk material.

What happens when the target is inside harvested cells?

Cells containing the target are disrupted mechanically, chemically, or enzymatically; the resulting debris is then removed.

What does process yield measure, and what can limit its value?

Yield is the amount of active product retained after a step or across the process. High recovery is not beneficial if it harms purity or product quality.

What is the purpose of the capture step?

Capture isolates and concentrates the target from clarified feed while removing a substantial portion of bulk impurities.

What does intermediate purification remove?

Intermediate purification removes remaining contaminant classes, such as host-cell proteins, residual DNA, or product variants.

What is the purpose of polishing?

Polishing targets specified trace impurities and unwanted product forms, such as aggregates or fragments.

What differences can chromatography use to separate a target from impurities?

Chromatography separates targets and impurities by differences such as binding affinity, electrical charge, size, or hydrophobicity.

Why does purification usually use multiple complementary steps?

No single operation removes every impurity, so a process combines complementary steps. Processing aids or leached chromatography ligands may also need control.

What do UF/DF operations do after purification?

Ultrafiltration and diafiltration (UF/DF) can concentrate the product and exchange it into a suitable formulation buffer.

What does product characterization establish?

Characterization establishes product identity and important attributes related to safety, purity, potency, and stability, such as structure, activity, and aggregation.