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Size-Exclusion Chromatography01:08

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In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
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Adsorption isotherms are mathematical models that describe how molecules in a gas or liquid phase interact with surfaces. Two of the most common isotherm models are the Langmuir and Freundlich isotherms, which relate to Type I monolayer chemisorption. The Langmuir model is based on four key assumptions:• Adsorption cannot exceed monolayer coverage.• All surface sites are equivalent.• Molecules adsorb only at vacant sites.• There are no interactions between adsorbed...
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Brunauer, Emmett, and Teller (BET) introduced a theory in 1938 that modified Langmuir's assumptions to explain multilayer physical adsorption. This theory is applicable to Type II isotherms and provides a more realistic picture of adsorption processes. The BET theory assumes a uniform solid surface with localized adsorption sites, where adsorption at one site doesn't affect adsorption at neighboring sites. This theory also allows for the possibility of additional molecules being adsorbed on top...
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Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
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Updated: Apr 15, 2026

Study of Short Peptide Adsorption on Solution Dispersed Inorganic Nanoparticles Using Depletion Method
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Thermo-responsive adsorbent for size-selective protein adsorption.

Micky Fu Xiang Lee1, Eng Seng Chan2, Kam Chiu Tam3

  • 1Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 46150 Bandar Sunway, Selangor, Malaysia.

Journal of Chromatography. A
|April 4, 2015
PubMed
Summary

This study developed a thermo-responsive adsorbent for size-selective protein separation. The poly(oligoethylene glycol) methacrylate (POEGMA) grafted onto cationized agarose effectively separated proteins based on size and temperature, enhancing recovery of larger biomolecules like Hepatitis B virus-like particles.

Keywords:
Ion exchange chromatographyLower critical solution temperature (LCST)Phase transitionPoly(oligoethylene glycol) methacrylate (POEGMA)Temperature-dependentThermo-responsive

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Area of Science:

  • Biomaterials Science
  • Separation Science
  • Polymer Chemistry

Background:

  • Developing selective adsorbents is crucial for biomolecule purification.
  • Thermo-responsive polymers offer tunable properties for separation applications.
  • Cationized agarose is a known adsorbent matrix for protein binding.

Purpose of the Study:

  • To investigate the use of POEGMA-grafted cationized agarose for size-selective protein adsorption.
  • To evaluate the impact of OEGMA content and temperature on protein adsorption.
  • To assess the adsorbent's performance in separating a mixture of biomolecules.

Main Methods:

  • Synthesis of cationized agarose grafted with varying contents of POEGMA.
  • Adsorption studies of bovine serum albumin (BSA) and myoglobin (Myo) at different temperatures.
  • Evaluation of a packed bed system for separating a protein mixture including Hepatitis B virus-like particle (HBVLP).

Main Results:

  • Increasing POEGMA content reduced BSA adsorption due to chain shielding.
  • Adsorption was temperature-dependent, with increased BSA adsorption above the polymer's lower critical solution temperature (LCST).
  • The adsorbent demonstrated enhanced recovery of HBVLP and selective flow-through of BSA at elevated temperatures.

Conclusions:

  • POEGMA-grafted cationized agarose exhibits thermo-responsive, size-selective protein adsorption capabilities.
  • The adsorbent shows potential for purifying large biomolecules by leveraging temperature-controlled conformational changes.
  • This material offers a promising platform for advanced bioseparation processes.