Immunity, inflammation and disease · 2024
Expression and Purification of ADAMTSL5 in E. coli and Validation of Activity in Psoriasis Serum.
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Qin Ru, Li Shangyang, Wu Boheng, Lin Ruilan, Yuan Yulin
Аннотация
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.
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.