Table 2. Typical limited forced degradation conditions.

Condition	Step	Duration	Tests
Low pH	Adjust pH with HCl to 3.0	48hr	UV, SEC, IEX, peptide mapping, potency
High pH	Adjust pH with NaOH to 9.0	48hr	UV, SEC, IEX, peptide mapping, potency
Light stress	1X ICH (UV, Visible)	N/A	UV, SEC, IEX, peptide mapping, potency
Methionine oxidation	0.05% tBHP	2–5 h	UV, SEC, IEX, peptide mapping, potency
pH jump	Coordinate with purification area	N/A	Measure A280, A320 and DLS before and after pH adjustment to pH5.5

tBHP, tert-Butyl hydroperoxide a reagent.⁵ ICH, International Conference on Harmonisation; details on guidance for Light stress can be found in ICH guideline Q1B.⁶ Details of the assays used during forced degradation are reported in Table S1