Hypothetical example - You have a formulation of 100 mg/mL protein, 20 mM histidine, 5% sucrose, 0.01% polysorbate-80, pH 6.2.

1. Calculate the osmolality...........mOsm/kg (assume protein does not contribute to

osmolality and histidine exists in acidic and basic forms in equal concentration). Report to the nearest whole number.

2. Calculate the Tg'............C. Report to nearest tenth of a degree (0.1).

3. Would an annealing step be appropriate for this product?

4. Assuming a 1 g fill weight, what is the required reconstitution volume?mL.

Report to the nearest thousandth of a mL (0.001). (Note, in this question we are filling by weight, so we don't need to worry about the density. If the question provided a fill volume, that would have to be converted to weight by using density).