Affinity

  • How Can Dyes Help with Chromatography?

How Can Dyes Help with Chromatography?

Nov 21 2018 Read 917 Times

There are many different types of chromatography, with new acronyms appearing regularly as researchers devise new approaches to improve existing methods and add new detectors or technology. But they all basically are reliant on what happens when the mobile and stationary phases interact. After all, chromatography is the prime separation science used in laboratories.

But researchers don’t just look to add high-tech and complex electronics to improve chromatography. Sometimes they are looking for a simple and more cost-effective solution. Affinity chromatography is one of the main chromatography techniques used to separate proteins in a mix. And one of the ways that researchers have ‘improved’ affinity chromatography is with the use of dyes.

Affinity chromatography – natural combination

Affinity chromatography is used to separate proteins based on the interaction between a protein - or protein group – and a ligand. The ligand is attached to the stationary phase or chromatography matrix. The key point is that the interaction between protein and ligand is reversible and can be controlled – by pH or other method – allowing different proteins to be held for different times. This allows the separation.

One of the unique points about affinity chromatography is that it allows purification of a biological molecule based on the molecules biological function or chemical structure. Affinity chromatography can separate biological mixtures that would otherwise be difficult to separate – offering high resolution, high selectivity and high capacity for the protein of choice.

The protocol for affinity chromatography is straightforward:

  • Load the column with the affinity ligands
  • Apply sample mix using a mobile phase – allow binding time.
  • Wash the column to remove non-bound biomolecules.
  • Elute required biomolecules using an appropriate mobile phase or by changing column condition.
  • Collect eluted sample.

Dye to be better?

One of the issues with affinity chromatography is that the ligands used to bind to the column matrices and attach to the proteins can be unstable, relatively insoluble and expensive. But it was found that synthetic dyes can be used as ligands for proteins. Synthetic dyes are relatively inexpensive, have a higher capacity than other ligands and are easily mobilized so they can be attached to the matrix and then removed.

The initial lack of specificity in dyes has been improved, leading to dyes known as biomimetic dyes. These mimic biological ligands allowing them to attach to the matrix and interact with the required proteins. Developments in chromatographic technology is discussed in the article, Column Technology for Achiral SFC Separations.

Reader comments

Do you like or dislike what you have read? Why not post a comment to tell others / the manufacturer and our Editor what you think. To leave comments please complete the form below. Providing the content is approved, your comment will be on screen in less than 24 hours. Leaving comments on product information and articles can assist with future editorial and article content. Post questions, thoughts or simply whether you like the content.

Post a Comment




Digital Edition

Chromatography Today - June 2019

June 2019

In This Edition Fundamental Aspects - High-resolution MS coupled to Photoionisation - Why do you Need to Measure BTEX in Ambient Air? - Gas Chromatography - Vacuum Ultraviolet Spectroscopy...

View all digital editions

Events

PREP 2019

Jul 07 2019 Baltimore, MD, USA

ISPPP 2019

Jul 10 2019 Baltimore, MD, USA

ACS National Meeting & Expo

Aug 25 2019 San Diego, CA, USA

Microscopy Conference 2019

Sep 01 2019 Berlin, Germany

BMSS Annual Meeting

Sep 03 2019 Manchester, UK

View all events