Preparation and characterization of monoclonal antibody against protein tyrosine phosphatase PRL-3
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Published on: May 18, 2021
Lirong Peng1, Yan Li, Ling Meng
1Department of Biochemistry and Molecular Biology, Beijing Institute for Cancer Research, Peking University School of Oncology, Beijing, People's Republic of China.
Abstract:
PRL-3 is a newly identified nonclassical protein tyrosine phosphatase associated with colorectal cancer metastasis. To prepare specific monoclonal antibody (MAb) against PRL-3, cDNAs of PRL-1, -2, and -3 were cloned by RT-PCR and inserted into prokaryotic expression vector pGEX-4T1, respectively. The fusion proteins were expressed in Escherichia coli. Monoclonal antibody against PRL-3 was obtained with hybridoma technique and specific ELISA screening. Western blotting and immunohistochemistry assays showed that MAb 3B6 had specific binding ability with PRL-3 protein in eukaryotic cells. This MAb will be a helpful tool for the detection of PRL-3 protein in the tissues and serum of colorectal cancer and other cancer patients.
Insights
Researchers developed a specific monoclonal antibody (MAb) to detect the PRL-3 protein, which is linked to colorectal cancer metastasis. This antibody offers a new tool for diagnosing colorectal cancer and other malignancies.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- PRL-3 is a nonclassical protein tyrosine phosphatase implicated in colorectal cancer metastasis.
- Targeting PRL-3 is crucial for developing new diagnostic and therapeutic strategies for cancer.
Purpose of the Study:
- To generate a specific monoclonal antibody (MAb) for detecting the PRL-3 protein.
- To validate the utility of the MAb in identifying PRL-3 in cancer tissues and serum.
Main Methods:
- Complementary DNAs (cDNAs) for PRL-1, -2, and -3 were cloned and expressed as fusion proteins in E. coli.
- Hybridoma technology and ELISA screening were employed to produce and select MAbs.
- Western blotting and immunohistochemistry were used to confirm the specificity and binding ability of MAb 3B6.
Main Results:
- A specific MAb, designated 3B6, was successfully generated against PRL-3.
- MAb 3B6 demonstrated specific binding to PRL-3 protein in eukaryotic cells.
- The antibody showed potential for detecting PRL-3 in colorectal cancer tissues and serum.
Conclusions:
- The developed MAb 3B6 is a valuable tool for detecting PRL-3.
- This antibody can aid in the diagnosis and monitoring of colorectal cancer and potentially other cancers associated with PRL-3.
