Scholarlink

<p>Recombinant protein expression using bacterial hosts has become crucial for mass-producing proteins, and effective purification is essential to ensure high yield and purity. However, the commonly used nickel-nitrilotriacetic acid (Ni-NTA) column chromatography has several limitations, including the necessity for cell lysate pretreatment, time-consuming processes, and restricted resin reusability. In this study, we synthesized two types of Ni-modified magnetic nanoparticles (MNPs)—Ni(II) oxide (NiO)-decorated zinc (Zn) ferrite nanoparticles (MNP1) and Ni-Zn ferrite nanoclusters (MNP2)—and evaluated their target protein binding capacities and ability to capture His<sub>6</sub>-amicyanin from soluble <i>Escherichia coli</i> extracts. Under purified protein conditions, MNP2 exhibited a higher apparent binding capacity than MNP1, likely due to its increased effective surface area. However, when applied to soluble <i>E. coli</i> periplasmic extracts, MNP1 achieved superior protein recovery, highlighting that intrinsic binding capacity does not necessarily translate to purification efficiency in complex biological environments. This performance reversal is attributed to differences in surface accessibility and colloidal stability between the two nanostructures. Both MNPs selectively captured His₆-amicyanin and maintained binding performance over multiple reuse cycles. Compared qualitatively with Ni-NTA chromatography, the MNP-based approach provided comparable recovery with a simplified workflow suitable for small-scale applications. These findings demonstrate that nanoparticle morphology and structural stability critically determine purification outcomes and provide design insights for developing structure-optimized magnetic systems for recombinant protein purification.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Purification of recombinant protein using magnetic NiO-decorated Zn ferrite nanoparticles and Ni-Zn ferrite nanoclusters as an alternative to Ni-NTA

  • 期刊论文

  • 发表日期：

    2026年4月30日

查看全文
  • Seulah Yang,
  • Eunjeong Kim,
  • Dahyeok Lee,
  • Yong Il Park,
  • Sooim Shin

摘要

Recombinant protein expression using bacterial hosts has become crucial for mass-producing proteins, and effective purification is essential to ensure high yield and purity. However, the commonly used nickel-nitrilotriacetic acid (Ni-NTA) column chromatography has several limitations, including the necessity for cell lysate pretreatment, time-consuming processes, and restricted resin reusability. In this study, we synthesized two types of Ni-modified magnetic nanoparticles (MNPs)—Ni(II) oxide (NiO)-decorated zinc (Zn) ferrite nanoparticles (MNP1) and Ni-Zn ferrite nanoclusters (MNP2)—and evaluated their target protein binding capacities and ability to capture His6-amicyanin from soluble Escherichia coli extracts. Under purified protein conditions, MNP2 exhibited a higher apparent binding capacity than MNP1, likely due to its increased effective surface area. However, when applied to soluble E. coli periplasmic extracts, MNP1 achieved superior protein recovery, highlighting that intrinsic binding capacity does not necessarily translate to purification efficiency in complex biological environments. This performance reversal is attributed to differences in surface accessibility and colloidal stability between the two nanostructures. Both MNPs selectively captured His₆-amicyanin and maintained binding performance over multiple reuse cycles. Compared qualitatively with Ni-NTA chromatography, the MNP-based approach provided comparable recovery with a simplified workflow suitable for small-scale applications. These findings demonstrate that nanoparticle morphology and structural stability critically determine purification outcomes and provide design insights for developing structure-optimized magnetic systems for recombinant protein purification.