FPLC in a Pipette Tip

Mar 12 2012

Author: Carole Staniford on behalf of Anachem

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The humble pipette tip has come of age. Now not just for transferring your sample, new functional tips actually perform your assay. Protein purification tips enable affinity purification that gives you higher concentration product with superior purity, faster and easier than ever before.

Many thousands of disposable pipette tips are used every day in laboratories worldwide. These simple plastic consumables may seem insignificant, but their quality plays a critical role in ensuring accurate and precise liquid dispensing. In addition to using a precision pipette that is regularly calibrated and being trained in correct pipetting technique, using pipette tips made in quality moulds from flexible virgin polypropylene is essential. The purity of the tip should also be considered as tips that are not BioClean could lead to contamination of samples or assay interference.
However, today the humble pipette tip is not just for transferring samples. Advances in technology have created a new generation of functional tips that actually contain and perform your assay. This new technique has significant advantages over traditional methods, for example for protein purification, providing  a superior end product in less time, yet remaining a low cost solution.

Protein Purification

A variety of methods are traditionally employed for purifying proteins and other macromolecules of interest from crude extracts or other complex mixtures. Most techniques, however, involve some form of chromatography whereby molecules in solution (mobile phase) are separated based
on differences in chemical or physical interaction with a stationary material (solid phase). Affinity chromatography (also called affinity purification) makes use of specific binding interactions between molecules. A particular ligand is chemically immobilised or ‘coupled’ to a solid support so that when a complex mixture is passed over the column, those molecules having specific binding affinity to the ligand become bound. After other sample components are washed away, the bound molecule is stripped from the support, resulting in its purification from the original sample.

A range of tools are currently available for affinity protein purification including: spin columns, gravity columns, FPLC and magnetic beads. Each specific affinity system requires its own set of conditions and presents its own peculiar challenges for a given research purpose. Protein Tips are novel protein capture devices using various affinity resins maintained at the base outlet of a pipette tip. They form part of a unique purification and enrichment system that includes an electronic pipette to automate and control the process of Capturing, Purifying and Enrichment of proteins. Protein Tips have been optimised to produce the highest possible purity, yield, concentration and activity of sample proteins. In addition, they are able to perform these functions with low volume samples, thus saving valuable resources. Sample work flow is seamlessly integrated with protein expression and assays, with complete sample and solution
tracking. The tips can operate rapidly in parallel to perform purification in minimal time, frequently in as little as 15 minutes. They are easy to set up and are disposable after use, making buffer preparation and clean up very easy.

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