Development and Characterization of PEGylated Chromatographic Monoliths as a Novel Platform for the Separation of PEGylated RNase a Isomers

Figure 3

Separation and observation of 40.0 kDa PEGylated RNase A isomers using the 20.0 kDa PEGylated monolith. (a) Chromatogram at 215 nm of 40.0 kDa PEGylated RNase A in the 20.0 kDa PEGylated monolith using a step gradient elution. Buffer A: Tris–HCl 20.0 mM pH 8.2. Buffer B : Buffer A + 3.0 M ammonium sulphate. Loop 0.1 mL, Volumetric flow 1 mL min⁻¹. (b) Silver staining for protein detection of SDS–PAGE analysis of the chromatographic fractions. (c) I₂-BaCl₂ staining for mPEG detection of SDS–PAGE analysis of chromatographic fractions.

(a)

(b)

(c)