PO0022 A?inity-based Dispersive Pipette Extraction for Automated Purification
來源:IMCS
|
作者:Todd Mullis1, Sunil Hwang1 ,Michael Mouradian2,L. Andrew Lee*1
|
發布時間: 959天前
|
928 次瀏覽
|
分享到:
A?nity-based extractions in complex matrixes are typically time-consuming due to the la-bor intensive work of hands on pipetting and centrifugation. A typical spin column a?nity extraction takes an hour to bind the sample and requires excess bu?er solutions towash o? non-speci?c binding proteins using multiple centrifugation steps. A quick, highthroughput a?nity-based enrichment method for target proteins is desired to allow forquicker screening of samples. We report a high-throughput sample preparation method us-ing antibody and his-tag based enrichment on a robotic liquid handling system with disper-sive pipette extraction. Speci?cally, Protein A and cobalt-IMAC resins were used for IgGenrichment from human serum and his-tagged proteins from cell lysates, respectively. Te Protein A extractions were compared side-by-side with a traditional spin-column based extraction protocol. Two di?erent his-tagged proteins were puri?ed fromcell lysate using the IMAC resins in tips. Tese developments allow for completely automat-ed 30 minute sample preparation leading to high-throughput sample screening and bettercompatibility with in-house automated liquid handling systems.
A?nity-based extractions in complex matrixes are typically time-consuming due to the la-bor intensive work of hands on pipetting and centrifugation. A typical spin column a?nity extraction takes an hour to bind the sample and requires excess bu?er solutions towash o? non-speci?c binding proteins using multiple centrifugation steps. A quick, highthroughput a?nity-based enrichment method for target proteins is desired to allow forquicker screening of samples. Herein, we report a high-throughput sample preparation method using antibody and his-tag based enrichment on a robotic liquid han-dling system with dispersive pipette extraction.
閱讀原文