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13Feb 2017

Affinity-based Dispersive Pipette Extraction for Automated Purification

affinity tips

ABSTRACT:

Affinity-based extractions in complex matrixes are typically time-consuming due to the labor intensive work of hands on pipetting and centrifugation. A typical spin column affinity extraction takes an hour to bind the sample and requires excess buffer solutions to wash off non-specific binding proteins using multiple centrifugation steps. A quick, high throughput affinity-based enrichment method for target proteins is desired to allow for quicker screening of samples. We report a high-throughput sample preparation method using antibody and his-tag based enrichment on a robotic liquid handling system with dispersive pipette extraction. Specifically, Protein A and CO-IMAC resins were used for IgG enrichment from human serum and his-tagged proteins from cell lysates, respectively. The Protein A extractions were compared side-by-side with a traditional spin-column based extraction protocol. Two different his-tagged proteins were purified from cell lysate using the CO-IMAC resins in tips. These developments allow for completely automated 30 minute sample preparation leading to high-throughput sample screening and better compatibility with in-house automated liquid handling systems.

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Download Application Note for affinity purification on theĀ Integra VIAFLO96

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