A Robust and Easy Protein Purification Method Using SpyDock-Modified Resin

Protein purification is a critical step in both life sciences and biomanufacturing. Traditional affinity chromatography (AC) methods, including His-tag-based purification, provide high-purity proteins but are limited by the high cost of resins and the need for additional tag-removal steps. In this p...

Full description

Saved in:
Bibliographic Details
Main Authors: Xiaofeng Yang, Zhanglin Lin, Ya Xiang, Binrui Chen, Zisha Lao
Format: Article
Language:English
Published: Bio-protocol LLC 2025-04-01
Series:Bio-Protocol
Online Access:https://bio-protocol.org/en/bpdetail?id=5270&type=0
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Protein purification is a critical step in both life sciences and biomanufacturing. Traditional affinity chromatography (AC) methods, including His-tag-based purification, provide high-purity proteins but are limited by the high cost of resins and the need for additional tag-removal steps. In this protocol, we present a reusable SpyDock-modified epoxy resin coupled with a pH-inducible self-cleaving intein for direct purification of proteins with authentic N-termini. This method enables efficient protein purification from cell lysates, achieving high purity (>90%) and yields comparable to the His-tag approach, without requiring tag removal. The SpyDock-modified resin protocol is robust, easy to implement, and cost-effective, making it suitable for both research and large-scale industrial applications.
ISSN:2331-8325