4th October 2019
Isolating a single target from a complex sample can be challenging and is often a multi step process which requires increased specificity for the target. Affinity separation is commonly used method as it involves the use of selective binding to purify proteins from a complex sample. Antibodies are commonly used as capture probes in tag-less purification method, however they come with their own limitations such as lengthy development process, sensitive to temperature and also undergo denaturation over the time. Aptamers offers the best alternative to antibodies for tag-less purification method as they are more stable and can be easily synthesized using our automated selection process under customer-defined conditions to yield highly specific aptamers. Moreover, due to their small size, aptamers can be more densely bound to a support, thereby increasing binding capacity.
Taking advantage of our highly specific aptamers, we have developed a process that enables the purification of tag-less proteins in their active state using aptamer mediated affinity chromatography (AMAC). Using this method, we have successfully purified a number of proteins from a complex mixture and eluted using customer-defined conditions (Figure 1).
Figure 1. Case study: SDS-PAGE analysis of a protein purified from expression medium using our highly specific aptamers. The protein was eluted using customerdefined buffer conditions for single step purification.
Aptamers isolated using our automated selection approach were readily immobilized on commercially available purification resin to give a target specific affinity resin. Customer defined wash and elution buffer conditions were used which gave a significantly purified product, without the risk associated with using antibody-based purification.
The sensitivity, precision and accuracy of the method were sufficient to purify proteins in their active state. This is mainly because of our customer defined selection process that enables to provide aptamers that are specific and have high affinity for the target. This method can be applied over a broad range of recombinant proteins and expression systems. If you would like to learn more about our selection strategies and purification process, please contact us using the form below.