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Before the sample solution was injected into the HPLC, extraction solution was injected into the HPLC sample bottle using a syringe containing a nylon membrane, which was used to protect the head from damage.
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In DLLME method, the extraction time is defined as the interval time between fast injecting a mixture of extraction and dispersive solvents into the sample solution before the start of centrifuges (Rezaee et al. 2006; Shamsipur and Fattahi 2011).
Then, 1.0 mL of phenolic solution (5%) and 5.0 mL of sulfuric acid were added to the sample solution before the absorbance was taken at 488 nm, spectrophotometrically (Shimadzu UV-1601, Japan).
The QB-mAbs were premixed with the sample solution before performing the test as a previously described protocol.
Before these chromatographic treatment procedures, the sample solution was passed through SPE cartridges.
Because diffusion-based separation is, in general, based on dilution of the sample solution, preconcentration before CE separation will be required in the next step.
The sample solution was filtered with a 0.22-µm filter before use.
This was the sample solution.
The final protein content in the sample solutions was quantified and then lyophilized before storage at −20°C.
As shown in the above photos of Figure 6, sample 1 is the original sample solution which was before the magnetic separation.
The samples were prepared such that the final sample solution contained 1× sample buffer and 1× NuPAGE sample reducing agent and were heated at 70°C for 10 min before loading.
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