Preparation of a novel monoclonal antibody against active components of PHA-L from Phaseolus vulgaris and its functional characteristics

• Published in: BMC biotechnology Vol. 22; no. 1; pp. 1 - 32
• Main Authors: Wang, Peipei, Hu, Junmei, Duan, Jiaqi, Min, Shitong, Chen, Congliang, Zhu, Yue, Pan, Yan, Wang, Yitian, Wei, Dapeng, Wang, Xia
• Format: Journal Article
• Language: English
• Published: London BioMed Central Ltd 29-10-2022; BioMed Central; BMC
• Subjects: AICD; Antigens; Apoptosis; Cancer; Care and treatment; Cell activation; Cell aggregation; Cell death; Cloning; Dosage and administration; Health aspects; Hybridoma clones; Immunofluorescence; Immunohistochemistry; Impurities; L protein; Lectins; Leukocytes; Lymphocytes; Lymphocytes T; Medical diagnosis; Molecular weight; Monoclonal antibodies; Monoclonal antibody; PHA-L; Plant lectins; Proteins; Tumors; China
• ISSN: 1472-6750; 1472-6750
• DOI: 10.1186/s12896-022-00761-7

Leukocyte phytohemagglutinin (PHA-L), derived from the L4 tetramer of PHA, has been frequently employed as a mitogen to induce T lymphocyte proliferation in vitro. The biological application of PHA-L in cancer diagnosis and treatment has gained traction in recent years. However, it has been noted that PHA-L obtained using traditional procedures has a massive amount of impurities or toxic components, which interfere with the activity of PHA-L. Preparation of a monoclonal antibody against active PHA-L is a significant tool for studying PHA-L's function and therapeutic potential. We successfully prepared monoclonal antibodies against the active components of PHA-L based on the whole PHA-L protein as an antigen, and found that monoclonal antibody 3C1C6G11 can be employed in western blot, immunofluorescence, and immunohistochemistry detection. Importantly, preliminary result shows that the mAb 3C1C6G11 may prevent PHA-L-induced cell aggregation and AICD (activation-induced cell death). The monoclonal antibody mAb 3C1C6G11 prepared in this study can be used as an effective tool for detecting PHA-L active components, investigating PHA-L's function and antineoplastic application.