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Dialysis01:15

Dialysis

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Dialysis is a diffusion-based purification process that separates analyte molecules from a complex matrix. This is accomplished by allowing molecules in the solution to pass through a semipermeable membrane into a liquid on the other side. The membrane is usually made of cellulose acetate or cellulose nitrate, and the second liquid must be miscible with the solution. Ions (e.g., chloride or sodium) or organic molecules (e.g., glucose) can pass through the membrane pores, which generally have...
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Extraction: Effects of pH00:53

Extraction: Effects of pH

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Consider a neutral form of an amine, B, with a partition coefficient, K, in a liquid mixture containing organic and aqueous phases. The pH of the aqueous phase affects the charge on acidic and basic solutes, and the charged form is usually more soluble in the aqueous phase. Suppose the conjugate acid form of the amine is soluble only in the aqueous phase while the base form is soluble in both phases. Then the distribution coefficient, D, can be given as the ratio of amine concentration in the...
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Protein Buffers in Blood Plasma and Cells01:20

Protein Buffers in Blood Plasma and Cells

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The human body utilizes protein buffer systems to maintain a stable pH. These systems capitalize on the dual role of amino acids, which can act as acids or bases by accepting or releasing hydrogen ions in response to pH changes. Protein buffer systems are particularly significant in the extracellular fluid (ECF) and intracellular fluid (ICF) of active cells, where structural and functional proteins provide substantial buffering capacity.
Certain amino acids can exist in a zwitterion state at a...
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Pore Transport and Ion-Pair Transport01:17

Pore Transport and Ion-Pair Transport

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Pore transport and ion-pair formation are critical mechanisms for the absorption and distribution of drugs in the body.
Pore transport, also known as convective transport, is a process where small molecules like urea, water, and sugars rapidly cross cell membranes as though there were channels or pores in the membrane. Although direct microscopic evidence is limited  but the concept of pores or channels is widely accepted based on physiological evidence. Despite the lack of direct...
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Patch Clamp01:18

Patch Clamp

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Many fundamental cell functions such as muscle contraction and nerve transmission rely on the electrical signals produced by the movement of positively and negatively charged ions across the cell membrane. One competent method to record current flowing across the whole cell or single ion channel is the patch-clamp technique.
In this method, a glass micropipette containing electrolyte solution is tightly sealed against a small portion of the cell membrane. As a result, a patch of the cell...
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SDS-PAGE01:27

SDS-PAGE

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Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed  polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact...
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Related Experiment Video

Updated: May 27, 2025

Sedimentation Equilibrium of a Small Oligomer-forming Membrane Protein: Effect of Histidine Protonation on Pentameric Stability
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Patchy Charge Distribution Affects the pH in Protein Solutions during Dialysis.

Sebastian P Pineda1, Pablo M Blanco2, Roman Staňo3,4

  • 1Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Hlavova 8, 128 40 Prague 2, Czech Republic.

Langmuir : the ACS Journal of Surfaces and Colloids
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Summary

Understanding pH differences during protein dialysis is crucial. This study introduces a model showing that patchy ionizable sites on proteins significantly impact pH, affecting purification outcomes.

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Area of Science:

  • Biophysical Chemistry
  • Protein Chemistry
  • Separation Science

Background:

  • Dialysis and diafiltration are used for protein purification, relying on buffer pH for product stability.
  • Achieving target pH in protein solutions during dialysis is challenging due to pH differences between permeate and retentate.
  • Existing models neglect the impact of protein surface charge distribution on pH.

Purpose of the Study:

  • To develop a computational model for understanding pH variations in protein solutions during dialysis.
  • To investigate the role of patchy distribution of ionizable sites on protein surface charge and pH.
  • To improve theoretical predictions for protein dialysis processes.

Main Methods:

  • A simple computational model of a colloidal particle with patchy acidic sites was developed.
  • Molecular simulations in the Grand-Reaction ensemble were employed.
  • Systematic variation of model parameters to analyze acid-base equilibria.

Main Results:

  • Interactions between ionizable sites significantly affect nanoparticle charge.
  • Patchy distribution of ionizable sites contributes to pH differences between permeate and retentate.
  • The effect of patchy distribution is more pronounced when sites are on smaller patches.

Conclusions:

  • Protein solutions exhibit similar physics to the model system.
  • Accurate prediction of pH during protein dialysis requires accounting for the patchy distribution of ionizable sites.
  • This finding is essential for optimizing protein purification processes and preventing product precipitation.