Affinity chromatography is a method of defining target proteins of a compound based on compound-target highly specific interactions. Creative Biolabs offers bioactive small molecules targets identification service for compounds. Targets are “fished” by immobilized compound and then recovered under mild conditions for purification and identification.

Affinity chromatography is a liquid chromatographic technique and it is widely used for purifying or separating biochemical mixtures based on highly specific interaction between two substances, e.g. antigen and antibody, enzyme and substrate, or receptor and ligand. In recent years, this technique has become increasingly important in defining the target proteins of a compound and understanding its mechanism of action.

The principle of affinity chromatography is simple. One of the interacting molecules referred to as affinity ligand, is immobilized onto a solid support, usually by covalent coupling, as “bait”. The target molecules are mixed in the mobile phase. When the mobile phase passes through immobilized solid support, the target molecule is captured by the “bait” in favorable conditions, while the other molecules with lower or lack of affinity to the ligand will directly pass through the column. The interactions between targets and hits are typically reversible. The sorbed compounds can be easily dissociated by changing the external conditions and washed. As a result, the target molecule is possible to be isolated, purified or gone through quantitative analysis.

Figure 1. Separation procedure in affinity chromatography (Acikara <em>et al.</em> 2013) Figure 1. Separation procedure in affinity chromatography (Acikara et al. 2013)

Creative Biolabs provides you affinity chromatography service to determine the binding affinity between targets and hits. We provide mild and efficient immobilization methods to preserve the molecular and conformational integrity and avoid your sensitive targets or compounds being destroyed. We choose the appropriate detergent, buffer, and protease inhibitors during the isolation procedure to minimize decreases due to denaturation, cleavage, enzymatic hydrolysis, etc. Our capabilities result in the desired specific enrichment of this technique while ensuring a high level of compatibility with downstream analysis by mass spectrometry.

Our affinity chromatography platform is especially suitable for target identification of natural therapeutic compounds. We prepare a customized affinity column in which the stationary phase is coated with the natural active compound. Cell lysate is acted as mobile phase and pass through the stationary phase to identify affinity analytes.

Identifying drug targets and knowing the mode of action (MOA) of a compound will increase the possibility of regulatory approval. Besides, potential toxicity caused by unwanted interaction between drugs and other functional proteins in human can also be identified to help researchers reduce the development risk of off-target interactions.

Creative Biolabs is proud to offer you affinity chromatography technique, which largely saving valuable drug development time. For more detailed information, please feel free to contact us or directly sent us an inquiry.

Reference

  1. Acikara Ö B, Çitoğlu G S, Özbilgin S et al. (2013). “Affinity Chromatography and Importance in Drug Discovery, In Column Chromatography, Chapter 3” InTech, DOI: 10.5772/55781.

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