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We saw that friction related effects were more important for small molecule separations whilst pressure effects were much more significant for protein separations. Insulin was an attractive example birli we could clearly see the decrease of retention in constant inlet pressure mode and the increase of retention in variable inlet pressure mode when increasing the flow rate.
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Therefore, we promote a generic method development approach which is based on the fact that proteins of the same, or similar, family (for example, mAbs or cys-linked ADCs) show very similar retention behavior. By knowing this, once you have developed a method for one mAb then it is possible to use these insights for another protein birli it will–elute with similar mobile phase strength, show good recovery only at high temperature (75 to 90 ºC), and only elute in a sharp peak when using an ion-pairing agent and low pH. Considering these facts, at the end you only need to perform very few experiments (in a more limited range of two or three method variables) bey input for further method optimization. The same input experiments güç be performed for any mAb – no need to study again the impact of method variables. Therefore, to optimize the method, retention modeling (computer simulation) hayat indeed be helpful.