N-GLYcanyzer monitoring of a microperfusion process reveals critical parameters' impact on Trastuzumab biosimilar N-glycoform time evolution

In a ~2 mL CHO microperfusion culture over 20 days, pH 6.7 and 39 {degrees}C drive Man5 accumulation and severe productivity penalties, while galactose plus MnCl2 shifts glycoforms toward G0F→G1F/G2F.

• Why it matters: Continuous intensified biomanufacturing needs dynamic control of mAb N-glycosylation, because end-point assays can miss time-evolving pathway responses that affect critical quality attributes.
• What they did: The team used N-GLYcanyzer PAT with offline assays and an integrated at-line workflow to screen six perfusion parameters, then tested feed supplementation with galactose and MnCl2 and an intensified perfusion rate of 1.2 VVD.
• The result: Nutrient limitation and thermal/acidic stress disrupted N-glycan maturation, but combining galactose/MnCl2 with 1.2 VVD sustained the galactosylation shift without a productivity cost, enabling attribute-based real-time control.

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