In the setup used in the case study (100 µL protein A column at 12 s residence time and Superdex 200 Increase 10/300), the biggest limitation lies in the 500 µl collected from the protein A column. When higher amounts were eluted from the protein A step, we lost protein as part of the peak was not collected. A solution would be to find a SEC column, onto which larger volumes can be injected, while keeping a good resolution. Increasing the residence time to, for example, 30 s will also increase the yield but triple the run time of the protein A step and consequently reduce the number of samples that can be processed. You also have to keep in mind that one should not load more than 1 mg on the Superdex 200 Increase 10/300 column.