Study of the mechanism of interaction of antibody (IgG) on two mixed mode sorbents

Ranjini, S. Shiva; Bimal, Devla; Dhivya, Ambur P.; Vijayalakshmi, M.A.

doi:10.1016/j.jchromb.2010.03.005

Cited by 20 publications

(9 citation statements)

References 17 publications

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“…7c), exhibiting a higher protein adsorption efficiency with mixedmode interactions at lower ISs and an excellent salt tolerance property in a wide IS range. The adsorption properties of γ-globulin on Bu60-PEI315 is comparable with and/or better than the results of other MMC resins (Du et al, 2014;Ranjini et al, 2010). For example, the γ-globulin adsorption capacity on CST-1 varied from 60 to 11 mg/mL at the NaCl concentration of 0.1-0.4 mol/L (Du et al, 2014).…”

Section: Protein Batch Uptakesupporting

confidence: 60%

Protein adsorption and chromatography on novel mixed-mode resins fabricated from butyl-modified poly(ethylenimine)-grafted Sepharose

Liu

Hong

et al. 2015

Chemical Engineering Science

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Section: Protein Batch Uptakesupporting

confidence: 60%

Protein adsorption and chromatography on novel mixed-mode resins fabricated from butyl-modified poly(ethylenimine)-grafted Sepharose

Liu

Hong

et al. 2015

Chemical Engineering Science

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“…Adsorption of proteins to hydrophobic ligand is entropy driven, which implies that the interaction increases with an increase in temperature (Hjerten, ). It has already been shown in a study with IgG as a model protein; the hydrophobic component is sensitive to increase in temperature and the presence of salt that leads to increased adsorption capacity of HEA (Ranjini et al ., ). It may be argued that increase in temperature might have led to few changes in the protein conformation thereby exposing more hydrophobic sites for interaction (Wu et al ., ).…”

Section: Discussionmentioning

confidence: 97%

“…These mixed‐mode ligands are recently gaining importance as the choice of sorbents for purification of many industrially important recombinant proteins (Brochier et al ., ; Cabanne et al ., ; Pezzini et al ., ), milk protein (Brochier et al ., ), polyclonal antibody (Ranjini et al ., ) and monoclonal antibody (Toueille et al ., ). All these reports show that these sorbents could be effectively used at the primary capture or intermediate purification step.…”

Section: Introductionmentioning

confidence: 99%

Study of catalase adsorption on two mixed‐mode ligands and the mechanism involved therein

Ranjini

Vijayalakshmi

2012

J of Molecular Recognition

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Mixed-mode chromatography sorbents n-hexylamine HyperCel™ (HEA) and phenylpropylamine HyperCel™ (PPA) were evaluated for the study of adsorption of catalase from two different sources. Various parameters such as buffer composition, ionic strength and pH were investigated to study the mechanism of interaction of commercially available pre-purified catalase from Bovine liver, purified catalase from black gram (Vigna mungo) and crude extract of black gram containing catalase with these mixed-mode ligands. A simple and economical screening protocol for identifying optimal buffer conditions for adsorption and desorption of catalase was established with micro volumes of the sorbent in batch mode. With HEA HyperCel, it was observed that pre-purified catalase from both bovine liver and black gram was completely retained at pH 7.0, irrespective of the presence or absence of NaCl in the adsorption buffer, whereas the catalase from crude extract of black gram was completely retained only in the presence of 0.2 M salt in the adsorption buffer. The elution of catalase from both the sources was accomplished by lowering the pH to 4.5 in absence of salt. In case of PPA HyperCel, catalase from both the sources was very strongly adsorbed under different buffer conditions studied, and elution did not yield a significant catalase activity. From the screening experiments, it could be concluded that the interaction of catalase with HEA HyperCel could be dominated by hydrophobic forces with minor contributions from ionic interaction and with PPA HyperCel, it could be a combination of different non-covalent interactions acting on different loci on the surface of the protein.

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“…However, they can also be seen as the basis for fine-tuning the separation of molecules that cannot be distinguished by the main mode of interaction alone. In fact, in recent years we have seen the introduction of a few dedicated mixed-mode stationary phases [1,2], mostly for the isolation of biopharmaceutical and diagnostic antibodies. Rekindled interest in hydroxyapatite, which is a natural mixed-mode material that has been known and used for the past six decades in bioseparation, is thus hardly surprising.…”

Section: Introductionmentioning

confidence: 99%

“…Hydroxyapatite (HA), with a stoichiometric formula of Ca 10 (PO 4 ) 6 (OH) 2 , is a calcium and phos-…”

Section: Introductionmentioning

confidence: 99%

Isolation and purification of recombinant proteins, antibodies and plasmid DNA with hydroxyapatite chromatography

Hilbrig

Freitag

2011

Biotechnology Journal

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Hydroxyapatite and related stationary phases increasingly play a role in the downstream processing of high-value biological materials, such as recombinant proteins, therapeutic antibodies and pharmaceutical-grade plasmid DNA. Chromatographic hydroxyapatite is an inorganic, ceramic material identical in composition, if not in structure, to calcium phosphate found in human bones and teeth. The interaction of hydroxyapatite with biomacromolecules is complex and highly dynamic, which can make predicting performance difficult, but also allows the design of very selective isolation processes. This review discusses the currently commercially available chromatographic materials, different retention mechanisms supported by these materials and differential exploitation for the design of highly specific isolation procedures. The state of the art of antibody purification by hydroxy-and fluoroapatite is reviewed together with tested routines for method development and implementation. Finally, the isolation of plasmid DNA is discussed, since the purification of DNA therapeutics at a sufficiently large scale is an emerging need in bioprocess development and perhaps the area in bioseparation where apatite chromatography can make its most important contribution to date.

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Study of the mechanism of interaction of antibody (IgG) on two mixed mode sorbents

Cited by 20 publications

References 17 publications

Protein adsorption and chromatography on novel mixed-mode resins fabricated from butyl-modified poly(ethylenimine)-grafted Sepharose

Protein adsorption and chromatography on novel mixed-mode resins fabricated from butyl-modified poly(ethylenimine)-grafted Sepharose

Study of catalase adsorption on two mixed‐mode ligands and the mechanism involved therein

Isolation and purification of recombinant proteins, antibodies and plasmid DNA with hydroxyapatite chromatography

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