Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials
Myoglobin (Mb) has been used as a biomarker for acute myocardial infarction. This study aimed to evaluate the stability of liquid Mb as quality control materials for Mb determination. Mb protein was expressed in Escherichia coli system and purified using Ni2+ chelate affinity chromatography. The pur...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-04-01
|
| Series: | Practical Laboratory Medicine |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352551725000095 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832557532369911808 |
|---|---|
| author | Yu-Hui Wang Xi-Feng Sun Chun-Xin Xu Feng-Qiang Sun Rong-Rong Wang Xiao-Kun Bian Zhan-Zhao Wang Qiang Wu |
| author_facet | Yu-Hui Wang Xi-Feng Sun Chun-Xin Xu Feng-Qiang Sun Rong-Rong Wang Xiao-Kun Bian Zhan-Zhao Wang Qiang Wu |
| author_sort | Yu-Hui Wang |
| collection | DOAJ |
| description | Myoglobin (Mb) has been used as a biomarker for acute myocardial infarction. This study aimed to evaluate the stability of liquid Mb as quality control materials for Mb determination. Mb protein was expressed in Escherichia coli system and purified using Ni2+ chelate affinity chromatography. The purity of purified recombinant Mb reached to 95 %. The immunoreactivity of Mb was investigated using Mb assay kits. The coefficient of determination (R2) of the curve fitted with dilution ratio and Mb concentration as variables was greater than 0.95, which indicated that Mb had good immunoreactivity. The concentrations per gradient measured using different kits had no significant difference (p-value>0.05), which indicated that the reactivity between the Mb antigen and Mb antibodies with different epitopes was good. The effects of different storage buffer, storage temperature and storage times on the stability of liquid Mb were investigated by detecting the concentration changes. At 2–8 °C for two months, Mb concentration in buffer B (Tris-HCl, pH 7.8, containing 1 % BSA and 0.05 % NaN3) decreased within 10 % compared with the initial concentration. The long-term storage stability was investigated by the thermal acceleration experiment. At 37 °C for one week, Mb concentration decreased by less than 15 %, indicating that the Mb had good long-term storage stability. The prepared liquid Mb had good immunoreactivity and stability, avoiding storage in freeze-dried powder. It was a promising alternative as the quality control material for Mb detection. |
| format | Article |
| id | doaj-art-98e1b7488c9f4bdcb1371e3a166a64d0 |
| institution | Kabale University |
| issn | 2352-5517 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Practical Laboratory Medicine |
| spelling | doaj-art-98e1b7488c9f4bdcb1371e3a166a64d02025-02-03T04:16:46ZengElsevierPractical Laboratory Medicine2352-55172025-04-0144e00456Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materialsYu-Hui Wang0Xi-Feng Sun1Chun-Xin Xu2Feng-Qiang Sun3Rong-Rong Wang4Xiao-Kun Bian5Zhan-Zhao Wang6Qiang Wu7Clinical Laboratory, Weifang People's Hospital, Shandong Second Medical University, Weifang, 261000, Shandong, ChinaClinical Laboratory, Weifang People's Hospital, Shandong Second Medical University, Weifang, 261000, Shandong, ChinaClinical Laboratory, Weifang People's Hospital, Shandong Second Medical University, Weifang, 261000, Shandong, ChinaClinical Laboratory, Weifang People's Hospital, Shandong Second Medical University, Weifang, 261000, Shandong, ChinaClinical Laboratory, Weifang People's Hospital, Shandong Second Medical University, Weifang, 261000, Shandong, ChinaClinical Laboratory, Weifang People's Hospital, Shandong Second Medical University, Weifang, 261000, Shandong, ChinaClinical Laboratory, Weifang People's Hospital, Shandong Second Medical University, Weifang, 261000, Shandong, ChinaCorresponding author.; Clinical Laboratory, Weifang People's Hospital, Shandong Second Medical University, Weifang, 261000, Shandong, ChinaMyoglobin (Mb) has been used as a biomarker for acute myocardial infarction. This study aimed to evaluate the stability of liquid Mb as quality control materials for Mb determination. Mb protein was expressed in Escherichia coli system and purified using Ni2+ chelate affinity chromatography. The purity of purified recombinant Mb reached to 95 %. The immunoreactivity of Mb was investigated using Mb assay kits. The coefficient of determination (R2) of the curve fitted with dilution ratio and Mb concentration as variables was greater than 0.95, which indicated that Mb had good immunoreactivity. The concentrations per gradient measured using different kits had no significant difference (p-value>0.05), which indicated that the reactivity between the Mb antigen and Mb antibodies with different epitopes was good. The effects of different storage buffer, storage temperature and storage times on the stability of liquid Mb were investigated by detecting the concentration changes. At 2–8 °C for two months, Mb concentration in buffer B (Tris-HCl, pH 7.8, containing 1 % BSA and 0.05 % NaN3) decreased within 10 % compared with the initial concentration. The long-term storage stability was investigated by the thermal acceleration experiment. At 37 °C for one week, Mb concentration decreased by less than 15 %, indicating that the Mb had good long-term storage stability. The prepared liquid Mb had good immunoreactivity and stability, avoiding storage in freeze-dried powder. It was a promising alternative as the quality control material for Mb detection.http://www.sciencedirect.com/science/article/pii/S2352551725000095Recombinant myoglobinQuality control materialEscherichia coli expression systemImmunoreactivityStorage stability |
| spellingShingle | Yu-Hui Wang Xi-Feng Sun Chun-Xin Xu Feng-Qiang Sun Rong-Rong Wang Xiao-Kun Bian Zhan-Zhao Wang Qiang Wu Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials Practical Laboratory Medicine Recombinant myoglobin Quality control material Escherichia coli expression system Immunoreactivity Storage stability |
| title | Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials |
| title_full | Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials |
| title_fullStr | Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials |
| title_full_unstemmed | Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials |
| title_short | Preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials |
| title_sort | preparation of recombinant myoglobin and investigation of the liquid antigen stability for quality control materials |
| topic | Recombinant myoglobin Quality control material Escherichia coli expression system Immunoreactivity Storage stability |
| url | http://www.sciencedirect.com/science/article/pii/S2352551725000095 |
| work_keys_str_mv | AT yuhuiwang preparationofrecombinantmyoglobinandinvestigationoftheliquidantigenstabilityforqualitycontrolmaterials AT xifengsun preparationofrecombinantmyoglobinandinvestigationoftheliquidantigenstabilityforqualitycontrolmaterials AT chunxinxu preparationofrecombinantmyoglobinandinvestigationoftheliquidantigenstabilityforqualitycontrolmaterials AT fengqiangsun preparationofrecombinantmyoglobinandinvestigationoftheliquidantigenstabilityforqualitycontrolmaterials AT rongrongwang preparationofrecombinantmyoglobinandinvestigationoftheliquidantigenstabilityforqualitycontrolmaterials AT xiaokunbian preparationofrecombinantmyoglobinandinvestigationoftheliquidantigenstabilityforqualitycontrolmaterials AT zhanzhaowang preparationofrecombinantmyoglobinandinvestigationoftheliquidantigenstabilityforqualitycontrolmaterials AT qiangwu preparationofrecombinantmyoglobinandinvestigationoftheliquidantigenstabilityforqualitycontrolmaterials |