In general, affinity chromatography consists of a stationary phase and a mobile phase. The mobile phase is a natural product lysate or any mixture containing biomolecules. The ligand that binds the target molecule is covalently attached to the solid phase. Affinity chromatography uses the interaction between the solid and mobile phases to obtain the pure substance you need. The target molecule binds to the ligand, while most other molecules flow through. The target biomolecule is eluted by changing conditions (pH or salt concentration) or by competing with the free ligand. Commonly used bioaffinities are enzyme-substrate, substrate analogs, exogenous lectin-polysaccharides, glycoproteins and other ion exchangers.
Affinity chromatography is generally very short, and care should be taken to select appropriate conditions when loading samples, including loading flow rate, buffer type, pH, ionic strength, temperature, etc., so that the substances to be separated can be fully bound to the affinity adsorbent.
Specific elution - As affinity adsorption is slow to reach equilibrium, specific elution often requires longer time and larger elution conditions, which can be eluted by changing other conditions, such as choosing substances with strong affinity.
Non-specific elution - When the substance to be separated is very strongly bound to the ligand, stronger acids or bases can be used or denaturing agents can be added to the eluent to deform the substance to be separated and dissociate it from the ligand.
BOC Sciences offers professional affinity chromatography services for the separation, identification and purification of components of mixtures of natural products for qualitative and quantitative analysis.
BOC Sciences offers a full range of integrated services for the isolation, extraction, purification and identification of natural products. Our services also include the customization of natural products and the optimization of affinity chromatography techniques to help our customers in the further analysis of natural products.
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