Binding and elution behavior of proteins on strong cation exchangers.

This work provides a broad survey of binding and elution behavior of proteins on strong cation exchangers. Four proteins comprising two monoclonal antibodies, lysozyme, and cytochrome c were used as models in the investigation. Seven chromatography resins with different base matrices were compared.

Comparison of agarose and dextran-grafted agarose strong ion exchangers for the separation of protein aggregates.

The control of aggregate levels in recombinant protein based drugs is a primary concern during process development and manufacture. In recent years, a novel class of dextran-grafted ion exchange matrices has gained popularity for process scale protein purification due to increased mass transfer rates and higher dynamic binding capacity compared to conventional matrices. Using bovine serum albumin and a monoclonal antibody as model proteins, we studied Sepharose FF and Sepharose XL ion exchangers for the separation of protein aggregates.

Comparison of strong anion-exchangers for the purification of a PEGylated protein.

We have studied the effect of protein PEGylation on ion-exchange adsorption using bovine serum albumin as a model system. The free sulfhydryl group of BSA, located on cysteine 34, was PEGylated using the maleimido-PEG chemistry. Several different BSA preparations were screened for extent of reaction using a 30 kDa PEG reagent.

Impact of denaturation with urea on recombinant apolipoprotein A-IMilano ion-exchange adsorption: equilibrium uptake behavior and protein mass transfer kinetics.

We have studied the equilibrium uptake behavior and mass transfer rate of recombinant apolipoprotein A-I(Milano) (apo A-I(M)) on Q Sepharose HP under non-denaturing, partially denaturing, and fully denaturing conditions. The protein of interest in this study is composed of amphipathic alpha helices that serve to solubilize and transport lipids. The dual nature of this molecule leads to the formation of micellar-like structures and self association in solution.