一种磷脂酶A2受体重组蛋白的制备方法
By expressing the protein in prokaryotic cells and combining it with freeze-thaw extraction and guanidine hydrochloride solution extraction, and using specific refolding solutions and multi-step purification techniques, the problem of low production efficiency of phospholipase A2 receptor recombinant protein was solved, and a highly active and immunoreactive recombinant protein was prepared, which is suitable for the detection of membranous nephropathy.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- DIYALAB ZJG BIOTECH CO LTD
- Filing Date
- 2022-11-22
- Publication Date
- 2026-07-17
AI Technical Summary
Existing technologies struggle to improve production efficiency while preserving the natural activity of recombinant phospholipase A2 receptor proteins. Furthermore, eukaryotic expression levels are low, and prokaryotic protein expression suffers from severe protein misfolding, making it difficult to meet the demands of large-scale production.
Recombinant phospholipase A2 receptor protein was expressed in prokaryotic cells. The protein was extracted using a combination of repeated freeze-thaw cycles and guanidine hydrochloride resuspension. The protein was then refolded using a specific refolding solution and purified using nickel ion affinity chromatography, cation exchange chromatography, and molecular sieve chromatography.
A high-yield, high-activity recombinant phospholipase A2 receptor protein was prepared, which can be recognized by antibodies in samples from patients with membranous nephritis. It exhibits superior immunoreactivity compared to the full-length PLA2R protein and has good storage stability, making it suitable for kits that detect antibodies or related diseases.
Smart Images
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