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Published on: May 18, 2021
Preparation and characterization of monoclonal antibody against protein tyrosine phosphatase PRL-3
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.
