Antibody purification: ion-exchange chromatography.

@article{Grodzki2010AntibodyPI,
  title={Antibody purification: ion-exchange chromatography.},
  author={Ana Cristina G. Grodzki and Elsa H. Berenstein},
  journal={Methods in molecular biology},
  year={2010},
  volume={588},
  pages={
          27-32
        }
}
Ion exchange chromatography techniques are the focus of this chapter and they showcase the power of this method for the purification of proteins and monoclonal antibodies. The technique is powerful and can separate biomolecules that have minor differences in their net charge, e.g., two protein molecules differing by a single charged amino acid. Given the amphoteric character of proteins the pH of the solution is important in the determination of the type of ion exchanger used. Immunoglobulins… Expand
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References

SHOWING 1-10 OF 133 REFERENCES
Rapid purification of monoclonal antibodies by high-performance liquid chromatography.
TLDR
Three different high-performance liquid chromatographic techniques have been used to purify monoclonal antibodies from ascites fluid and a combination of ion-exchange and hydrophobic interaction chromatography resulted in an antibody preparation comparable in purity to the one obtained from ammonium sulphate-precipitated immunoglobulin. Expand
Factorial screening of antibody purification processes using three chromatography steps without protein A.
Protein A affinity chromatography is often employed as a capture step to meet the purity, yield, and throughput requirements for pharmaceutical antibody purification. However, a trade-off existsExpand
Antibody separation by hydrophobic charge induction chromatography.
TLDR
Hydrophobic charge induction chromatography using 4-mercapto-ethyl-pyridine as the ligand is an effective method for the separation of antibodies from a variety of feedstocks and can reach levels of purity as high as 98%. Expand
Comparison of hydrophobic charge induction chromatography with affinity chromatography on protein A for harvest and purification of antibodies.
TLDR
P pH-based control of chromatographic function was demonstrated, and Chromatography on this antibody-selective HCIC sorbent was evaluated as a cost-effective, process-compatible alternative to affinity chromatography protein A sorbents. Expand
[26] Chromatography of proteins on ion-exchange adsorbents
Publisher Summary This chapter outlines general considerations which govern the use of ion-exchange chromatographic experiments emphasizing the precautions necessitated by the use of commercialExpand
Adsorption of monoclonal antibody variants on analytical cation-exchange resin.
TLDR
A lumped kinetic model and a procedure for estimating the corresponding parameters were developed and validated by comparison with experimental elution chromatograms in a wide range of operating conditions, and corresponding optimal values of pH and salt concentration were determined. Expand
Purification of monoclonal antibodies from ascites using ABx liquid chromatography column.
Murine and rat monoclonal antibodies (MAbs) were purified to 90-95% homogeneity from ascites in a single step using an ABx liquid chromatography column which utilizes a silica-based mixed-modeExpand
New method for the selective capture of antibodies under physiolgical conditions
TLDR
One example of antibody purification process involving hydrophobic charge induction chromatography as a capture step followed by a polishing phase with DEAE Ceramic HyperD is described, finding longevity and ligand leakage are compatible with large-scale applications. Expand
Aspects of Modeling a preparative ion-exchange step for antibody purification
In protein chromatography large molecules have to diffuse into the stationary phase. The molecular sizes of the proteins make the diffusion rates low or very low and the separation can be assumed toExpand
Cation exchange chromatography in antibody purification: pH screening for optimised binding and HCP removal.
TLDR
It is demonstrated with the principle of residence time that scout columns are suitable for dynamic studies and the optimum pH range from batch binding was transferred to scout columns which were then used to screen for maximum dynamic capacities, and IEF titration curve analysis was employed to define a final operational pH. Expand
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