Chromatography thiophilic

Table 4.8. Binding affinities of immunoglobulin subclasses from different species to immobilized Protein A, Protein G, Protein L, and thiophilic chromatography supports...

Thiophilic chromatography depends on the binding of proteins to sulphur atoms contained in a particular chemical context. During the development of methods for the preparation of affinity supports... 

Boschetti, E. (2001) The use of thiophilic chromatography for antibody purification a review. J. Biochem. Biophys. Methods. 49, 361-389. 

Thiophilic chromatography for the separation of antibody was also applied to the extraction of IgG from sweet cheese whey. The purity of immunoglobulin G was about 75% after a single separation and required only the addition of sodium or potassium sulfate to the initial feedstock.156... 

Contrary to thiophilic chromatography, HCIC does not require the presence of salts during the adsorption phase, thus immunoglobulin capture can be effected in physiological conditions and pH close to neutrality no feedstock pretreatment is required. Figure 17 shows no effect of ionic... 

Other methods that are related to affinity chromatography include hydrophobic interaction chromatography and thiophilic adsorption. The former is based on the interactions of proteins, peptides, and nucleic acids with short nonpolar chains on a support. This was first described in 1972 [113,114] following work that examined the role of spacer arms on the nonspecific adsorption of affinity columns [114]. Thiophilic adsorption, also known as covalent or chemisorption chromatography, makes use of immobilized thiol groups for solute retention [115]. Applications of this method include the analysis of sulfhydryl-containing peptides or proteins and mercurated polynucleotides [116]. 

Thiophilic adsorption chromatography has been described for the purification of murine monoclonal antibodies from hybridoma cell culture containing fetal bovine serum.154 Due to the very low concentration of immunoglobulins in cell culture supernatants, binding capacity remains modest in spite of the presence of 0.5 to 1 M potassium sulfate. Further developments of this technology described by Nopper et al.155 indicated that thiophilic sorbents could be modified in their structure to increase the specificity and the binding capacity. 

Nopper, B., Kohen, F., and Wilchek, M. (1989). A thiophilic adsorbent for the one-step high-performance liquid chromatography purification of monoclonal antibodies. Anal. Biochem. 180, 66-71. 

Knudsen, K. L., Hansen, M. B., Henriksen, L. R., Andersen, B. K., and Lihme, A. (1992). Sulfone-aromatic ligands for thiophilic adsorption chromatography Purification of human and mouse immunoglobulins. Anal. Biochem. 201, 170-177. 

Schwarz A. (1996). Five-membered mercaptoheterocyclic ligands for thiophilic adsorption chromatography. J. Mol. Recognition 6, 672-674. 

Hansen, P., Scobie, J. A., Hanson, B., and Hoogenraad, N. J. (1998). Isolation and purification of immunoglobulins from chicken eggs using thiophilic interaction chromatography. J. Immunol Methods 215, 1-7. 

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