Thermal unfolding of a lot of proteins can be partly reversible as long as the temperature is not increased beyond a critical value that induces non-reversible changes in the protein structure. Finding this temperature requires a different heating program than the widely used linear temperature ramp that is commonly applied to measure T_m values. In this protocol, the temperature of non-reversible unfolding of Trastuzumab (Herceptin) is determined using the Incremental Cycle mode of the PR.TimeControl software. Herceptin is a monoclonal antibody used to treat breast cancer.

thermal cycling | refolding | non-reversibility curve | PR-P-018