Establishment of matrix metalloproteinase 3 time-resolved immunoassay and some potential clinical applications
To develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum matrix metalloproteinase-3 (MMP-3) and to assess MMP-3's clinical value in patients with colorectal cancer (CRC).st. MMP-3 levels were established using the double antibody sandwich technique...
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Published in | Analytical biochemistry Vol. 666; p. 115072 |
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Main Authors | , , , , , , , , , , , |
Format | Journal Article |
Language | English |
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United States
Elsevier Inc
01.04.2023
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ISSN | 0003-2697 1096-0309 1096-0309 |
DOI | 10.1016/j.ab.2023.115072 |
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Abstract | To develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum matrix metalloproteinase-3 (MMP-3) and to assess MMP-3's clinical value in patients with colorectal cancer (CRC).st.
MMP-3 levels were established using the double antibody sandwich technique. The MMP-3 TRFIA technique was developed and optimized, and its linearity, sensitivity, accuracy, specificity, and recovery were assessed. Then, serum concentrations in healthy individuals and patients with CRC were determined by MMP-3 TRFIA.
The linear range of MMP-3 TRFIA was 0.73–500 ng/mL. MMP-3 TRFIA had an intra-batch precision range of 2.16%–7.10% percent and an inter-batch precision range of 3.99%–11.21%. MMP-3, tumor-associated trypsinogen 2, and AFP had no cross reaction.The recovery is between 90% and 110%, and had no serum interference. Patients with CRC had serum MMP-3 levels (73.95 ± 78.43 ng/mL) that were considerably higher than those of healthy individuals (21.45 ± 11.12 ng/mL), and those with metastasis had serum MMP-3 levels (95.89 ± 76.21 ng/mL) that were considerably higher than those of patients without metastasis (52.74 ± 47.25 ng/mL).
A highly sensitive MMP-3 TRFIA assay was successfully developed, and serum MMP-3 may be associated with CRC invasion and metastasis. Therefore, MMP-3 can be used in the auxiliary diagnosis of CRC.
•A time-resolved fluorescence immunoassay method for the detection of serum MMP-3 concentration was successfully established.•We found the levels of serum MMP-3 were higher in patients with colorectal cancer than healthy subjects.•Detection of serum MMP-3 concentration can distinguish patients with and without metastatic colorectal cancer.•MMP-3 can be used as an auxiliary detection marker for colorectal cancer. |
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AbstractList | To develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum matrix metalloproteinase-3 (MMP-3) and to assess MMP-3's clinical value in patients with colorectal cancer (CRC).st.AIMTo develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum matrix metalloproteinase-3 (MMP-3) and to assess MMP-3's clinical value in patients with colorectal cancer (CRC).st.MMP-3 levels were established using the double antibody sandwich technique. The MMP-3 TRFIA technique was developed and optimized, and its linearity, sensitivity, accuracy, specificity, and recovery were assessed. Then, serum concentrations in healthy individuals and patients with CRC were determined by MMP-3 TRFIA.METHODSMMP-3 levels were established using the double antibody sandwich technique. The MMP-3 TRFIA technique was developed and optimized, and its linearity, sensitivity, accuracy, specificity, and recovery were assessed. Then, serum concentrations in healthy individuals and patients with CRC were determined by MMP-3 TRFIA.The linear range of MMP-3 TRFIA was 0.73-500 ng/mL. MMP-3 TRFIA had an intra-batch precision range of 2.16%-7.10% percent and an inter-batch precision range of 3.99%-11.21%. MMP-3, tumor-associated trypsinogen 2, and AFP had no cross reaction.The recovery is between 90% and 110%, and had no serum interference. Patients with CRC had serum MMP-3 levels (73.95 ± 78.43 ng/mL) that were considerably higher than those of healthy individuals (21.45 ± 11.12 ng/mL), and those with metastasis had serum MMP-3 levels (95.89 ± 76.21 ng/mL) that were considerably higher than those of patients without metastasis (52.74 ± 47.25 ng/mL).RESULTSThe linear range of MMP-3 TRFIA was 0.73-500 ng/mL. MMP-3 TRFIA had an intra-batch precision range of 2.16%-7.10% percent and an inter-batch precision range of 3.99%-11.21%. MMP-3, tumor-associated trypsinogen 2, and AFP had no cross reaction.The recovery is between 90% and 110%, and had no serum interference. Patients with CRC had serum MMP-3 levels (73.95 ± 78.43 ng/mL) that were considerably higher than those of healthy individuals (21.45 ± 11.12 ng/mL), and those with metastasis had serum MMP-3 levels (95.89 ± 76.21 ng/mL) that were considerably higher than those of patients without metastasis (52.74 ± 47.25 ng/mL).A highly sensitive MMP-3 TRFIA assay was successfully developed, and serum MMP-3 may be associated with CRC invasion and metastasis. Therefore, MMP-3 can be used in the auxiliary diagnosis of CRC.CONCLUSIONSA highly sensitive MMP-3 TRFIA assay was successfully developed, and serum MMP-3 may be associated with CRC invasion and metastasis. Therefore, MMP-3 can be used in the auxiliary diagnosis of CRC. To develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum matrix metalloproteinase-3 (MMP-3) and to assess MMP-3's clinical value in patients with colorectal cancer (CRC).st. MMP-3 levels were established using the double antibody sandwich technique. The MMP-3 TRFIA technique was developed and optimized, and its linearity, sensitivity, accuracy, specificity, and recovery were assessed. Then, serum concentrations in healthy individuals and patients with CRC were determined by MMP-3 TRFIA. The linear range of MMP-3 TRFIA was 0.73–500 ng/mL. MMP-3 TRFIA had an intra-batch precision range of 2.16%–7.10% percent and an inter-batch precision range of 3.99%–11.21%. MMP-3, tumor-associated trypsinogen 2, and AFP had no cross reaction.The recovery is between 90% and 110%, and had no serum interference. Patients with CRC had serum MMP-3 levels (73.95 ± 78.43 ng/mL) that were considerably higher than those of healthy individuals (21.45 ± 11.12 ng/mL), and those with metastasis had serum MMP-3 levels (95.89 ± 76.21 ng/mL) that were considerably higher than those of patients without metastasis (52.74 ± 47.25 ng/mL). A highly sensitive MMP-3 TRFIA assay was successfully developed, and serum MMP-3 may be associated with CRC invasion and metastasis. Therefore, MMP-3 can be used in the auxiliary diagnosis of CRC. To develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum matrix metalloproteinase-3 (MMP-3) and to assess MMP-3's clinical value in patients with colorectal cancer (CRC).st. MMP-3 levels were established using the double antibody sandwich technique. The MMP-3 TRFIA technique was developed and optimized, and its linearity, sensitivity, accuracy, specificity, and recovery were assessed. Then, serum concentrations in healthy individuals and patients with CRC were determined by MMP-3 TRFIA. The linear range of MMP-3 TRFIA was 0.73-500 ng/mL. MMP-3 TRFIA had an intra-batch precision range of 2.16%-7.10% percent and an inter-batch precision range of 3.99%-11.21%. MMP-3, tumor-associated trypsinogen 2, and AFP had no cross reaction.The recovery is between 90% and 110%, and had no serum interference. Patients with CRC had serum MMP-3 levels (73.95 ± 78.43 ng/mL) that were considerably higher than those of healthy individuals (21.45 ± 11.12 ng/mL), and those with metastasis had serum MMP-3 levels (95.89 ± 76.21 ng/mL) that were considerably higher than those of patients without metastasis (52.74 ± 47.25 ng/mL). A highly sensitive MMP-3 TRFIA assay was successfully developed, and serum MMP-3 may be associated with CRC invasion and metastasis. Therefore, MMP-3 can be used in the auxiliary diagnosis of CRC. To develop a highly sensitive time-resolved fluorescence immunoassay (TRFIA) for the detection of serum matrix metalloproteinase-3 (MMP-3) and to assess MMP-3's clinical value in patients with colorectal cancer (CRC).st. MMP-3 levels were established using the double antibody sandwich technique. The MMP-3 TRFIA technique was developed and optimized, and its linearity, sensitivity, accuracy, specificity, and recovery were assessed. Then, serum concentrations in healthy individuals and patients with CRC were determined by MMP-3 TRFIA. The linear range of MMP-3 TRFIA was 0.73–500 ng/mL. MMP-3 TRFIA had an intra-batch precision range of 2.16%–7.10% percent and an inter-batch precision range of 3.99%–11.21%. MMP-3, tumor-associated trypsinogen 2, and AFP had no cross reaction.The recovery is between 90% and 110%, and had no serum interference. Patients with CRC had serum MMP-3 levels (73.95 ± 78.43 ng/mL) that were considerably higher than those of healthy individuals (21.45 ± 11.12 ng/mL), and those with metastasis had serum MMP-3 levels (95.89 ± 76.21 ng/mL) that were considerably higher than those of patients without metastasis (52.74 ± 47.25 ng/mL). A highly sensitive MMP-3 TRFIA assay was successfully developed, and serum MMP-3 may be associated with CRC invasion and metastasis. Therefore, MMP-3 can be used in the auxiliary diagnosis of CRC. •A time-resolved fluorescence immunoassay method for the detection of serum MMP-3 concentration was successfully established.•We found the levels of serum MMP-3 were higher in patients with colorectal cancer than healthy subjects.•Detection of serum MMP-3 concentration can distinguish patients with and without metastatic colorectal cancer.•MMP-3 can be used as an auxiliary detection marker for colorectal cancer. |
ArticleNumber | 115072 |
Author | Qin, Yuan Fang, Hongming Fu, Yulin Hong, Jianfeng Chen, Xindong Zhou, Xiumei Wang, Yigang Zhou, Zixuan Wang, Xiaoyan Zhou, Jingnan Huang, Biao Liu, Pengfei |
Author_xml | – sequence: 1 givenname: Yulin surname: Fu fullname: Fu, Yulin organization: College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China – sequence: 2 givenname: Xiaoyan surname: Wang fullname: Wang, Xiaoyan organization: Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, China – sequence: 3 givenname: Xindong surname: Chen fullname: Chen, Xindong organization: College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China – sequence: 4 givenname: Jianfeng surname: Hong fullname: Hong, Jianfeng organization: Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, China – sequence: 5 givenname: Yuan surname: Qin fullname: Qin, Yuan organization: College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China – sequence: 6 givenname: Zixuan surname: Zhou fullname: Zhou, Zixuan organization: College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China – sequence: 7 givenname: Xiumei surname: Zhou fullname: Zhou, Xiumei organization: College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China – sequence: 8 givenname: Yigang surname: Wang fullname: Wang, Yigang organization: College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China – sequence: 9 givenname: Jingnan surname: Zhou fullname: Zhou, Jingnan organization: Zhejiang Cancer Hospital, Hangzhou, China – sequence: 10 givenname: Hongming surname: Fang fullname: Fang, Hongming organization: Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, China – sequence: 11 givenname: Pengfei surname: Liu fullname: Liu, Pengfei organization: The Jiangyin Clinical College of Xuzhou Medical University, Jiangyin, China – sequence: 12 givenname: Biao orcidid: 0000-0003-1883-0670 surname: Huang fullname: Huang, Biao email: jswxhb@163.com organization: College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China |
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Keywords | Diagnosis Double antibody sandwich TRFIA MMP-3 Colorectal cancer |
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SubjectTerms | Antibodies blood serum Colorectal cancer colorectal neoplasms Diagnosis Double antibody sandwich fluorescent antibody technique Fluoroimmunoassay - methods Humans immunoassays Matrix Metalloproteinase 3 metastasis MMP-3 Serum stromelysin 1 TRFIA trypsinogen |
Title | Establishment of matrix metalloproteinase 3 time-resolved immunoassay and some potential clinical applications |
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