Expression and Purification of ADAMTSL5 in E. coli and Validation of Activity in Psoriasis Serum

IF 3.1 4区医学 Q3 IMMUNOLOGY

Immunity, Inflammation and Disease Pub Date : 2024-12-13 DOI:10.1002/iid3.70100

Ru Qin, Shangyang Li, Boheng Wu, Ruilan Lin, Yulin Yuan

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引用次数: 0

Abstract

Purpose

A disintegrin and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5), a protein linked to psoriasis, was obtained by prokaryotic expression and purification for potential utilization as a new auxiliary diagnostic marker for psoriasis. It was subsequently applied in psoriasis research.

Experimental Design

The designed ADAMTSL5 gene was inserted into the pET-30a (+) vector and expressed in the BL21 (DE3) strain as a fusion protein. Following this, the recombinant ADAMTSL5 protein was purified using affinity chromatography. Purified ADAMTSL5, obtained in conjunction with magnetic beads, was directly employed in both psoriasis patients and healthy individuals.

Results

The results indicated that the molecular weight of the ADAMTSL5 protein obtained through prokaryotic expression and purification was approximately 27 kDa, with a protein concentration of 0.96 mg/mL. Analysis of anti-ADAMTSL5 levels in the serum of both psoriasis patients and healthy individuals using magnetic microparticle chemiluminescence demonstrated that anti-ADAMTSL5 levels in the serum of psoriasis patients surpassed those of healthy individuals, showing a significant difference with p < 0.001.

Conclusions

The experimental findings presented here may contribute to the utilization of ADAMTSL5 as a marker for diagnosing psoriasis, offering novel insights and experimental avenues for further investigation.

Abstract Image

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ADAMTSL5在大肠杆菌中的表达纯化及银屑病血清活性验证

目的：通过原核表达和纯化获得银屑病相关蛋白ADAMTSL5 (disintegrin and metalloprotease domain containing thrombospondin type 1 motif-like 5)，有望作为银屑病新的辅助诊断标志物。它随后被应用于牛皮癣的研究。实验设计：将设计的ADAMTSL5基因插入pET-30a（+）载体中，以融合蛋白的形式在BL21 （DE3）菌株中表达。随后，利用亲和层析纯化重组ADAMTSL5蛋白。纯化的ADAMTSL5与磁珠一起获得，直接用于牛皮癣患者和健康个体。结果：经原核表达纯化得到的ADAMTSL5蛋白分子量约为27 kDa，蛋白浓度为0.96 mg/mL。磁微粒化学发光法分析银屑病患者和健康人血清中抗adamtsl5水平，银屑病患者血清中抗adamtsl5水平高于健康人，与p有显著性差异。本文的实验结果可能有助于将ADAMTSL5作为牛皮癣的诊断标志物，为进一步的研究提供新的见解和实验途径。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Immunity, Inflammation and Disease Medicine-Immunology and Allergy

CiteScore

3.60

自引率

0.00%

发文量

146

审稿时长

8 weeks

期刊介绍： Immunity, Inflammation and Disease is a peer-reviewed, open access, interdisciplinary journal providing rapid publication of research across the broad field of immunology. Immunity, Inflammation and Disease gives rapid consideration to papers in all areas of clinical and basic research. The journal is indexed in Medline and the Science Citation Index Expanded (part of Web of Science), among others. It welcomes original work that enhances the understanding of immunology in areas including: • cellular and molecular immunology • clinical immunology • allergy • immunochemistry • immunogenetics • immune signalling • immune development • imaging • mathematical modelling • autoimmunity • transplantation immunology • cancer immunology