Protein adsorption and transport in agarose and dextran-grafted agarose media for ion exchange chromatography

2007 ◽  
Vol 1146 (2) ◽  
pp. 202-215 ◽  
Author(s):  
Melani C. Stone ◽  
Giorgio Carta
Keyword(s):  
Ion Exchange ◽  
Protein Adsorption ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography

scholarly journals Isothermal Titration Calorimetry as a Method for Analyzing Protein Adsorption in Ion Exchange Chromatography

2021 ◽  
Vol 333 ◽  
pp. 15001
Author(s):  
Joao Simoes-Cardoso ◽  
Nanako Hoshino ◽  
Noriko Yoshimoto ◽  
Shuichi Yamamoto
Keyword(s):  
Ion Exchange ◽  
Protein Adsorption ◽  
Isothermal Titration Calorimetry ◽  
Protein Interactions ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Titration Calorimetry ◽  
Valuable Insight ◽  
Adsorption Enthalpy ◽  
Binding Behavior

Ion exchange chromatography is a widely used method for purification of all types of biomolecules in current biotechnological downstream processes. Knowledge on the binding behavior of proteins provides valuable insight for understanding the molecular mechanisms of protein interactions in a biological context. However, thermodynamic parameters such as enthalpy and entropy changes that characterize protein adsorption are still unknown. Isothermal titration calorimetry applications in biosciences has gained its merit to study binding of soluble molecules, protein inhibition, conformational changes, reaction kinetics, and protein adsorption. However, in the case of protein adsorption, a lot of complications arise since the usual models used to study protein interactions in solution are no longer valid. This work explains a detailed methodology for the obtention of adsorption enthalpy, entropy and Gibbs energy of protein adsorption, by using ITC together with equilibrium adsorption isotherms.

Studies on an Inhibitor of Plasminogen Activation in Human Serum

1973 ◽  
Vol 30 (02) ◽  
pp. 414-424 ◽  
Author(s):  
Ulla Hedner
Keyword(s):  
Ion Exchange ◽  
Human Serum ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Specific Adsorption ◽  
Partial Purification ◽  
Plasminogen Activation ◽  
Preparative Electrophoresis

SummaryA procedure is described for partial purification of an inhibitor of the activation of plasminogen by urokinase and streptokinase. The method involves specific adsorption of contammants, ion-exchange chromatography on DEAE-Sephadex, gel filtration on Sephadex G-200 and preparative electrophoresis. The inhibitor fraction contained no antiplasmin, no plasminogen, no α1-antitrypsin, no antithrombin-III and was shown not to be α2 M or inter-α-inhibitor. It contained traces of prothrombin and cerulo-plasmin. An antiserum against the inhibitor fraction capable of neutralising the inhibitor in serum was raised in rabbits.

Review on the Application of Mixed-mode Chromatography for Separation of Structure Isoforms

2018 ◽  
Vol 20 (1) ◽  
pp. 56-60 ◽  
Author(s):  
Tsutomu Arakawa
Keyword(s):  
Ion Exchange ◽  
Mixed Mode ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Charged State ◽  
Partial Structure ◽  
Reverse Phase ◽  
Oligomer Formation ◽  
Structure Rearrangement ◽  
Disulfide Scrambling

Proteins often generate structure isoforms naturally or artificially due to, for example, different glycosylation, disulfide scrambling, partial structure rearrangement, oligomer formation or chemical modification. The isoform formations are normally accompanied by alterations in charged state or hydrophobicity. Thus, isoforms can be fractionated by reverse-phase, hydrophobic interaction or ion exchange chromatography. We have applied mixed-mode chromatography for fractionation of isoforms for several model proteins and observed that cation exchange Capto MMC and anion exchange Capto adhere columns are effective in separating conformational isoforms and self-associated oligomers.

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