Preparation of monoclonal antibodies against human leukocyte immunoglobulin like receptor B2 and preliminary identification of their activity in vitro
10.13200/j.cnki.cjb.004483
- VernacularTitle:抗人白细胞免疫球蛋白样受体亚家族B2单克隆抗体的制备及体外活性初步鉴定
- Author:
FENG Yanping,HU
- Publication Type:Journal Article
- Keywords:
Leukocyte immunoglobulin like receptor subfamily B2(LILRB2);
Tumor immunity;
Immune checkpoint;
Monoclonal antibodies
- From:
Chinese Journal of Biologicals
2025;38(05):549-556
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare monoclonal antibodies against human leukocyte immunoglobulin like receptor subfamilyB2(LILRB2) by hybridoma technique and preliminarily identify their activity in vitro, so as to provide a new strategy for targeting“cold”tumor therapy.MethodsThe extracellular region of human LILRB2 protein was expressed, purified by affinity chro-matography, and then used to immunize BALB/c mice to prepare anti-LILRB2 monoclonal antibodies with high specificityand affinity. The variable regions of light chain and heavy chain of mouse monoclonal antibodies were inserted into an expres-sion vector containing human constant regions by DNA recombination technique to produce chimeric antibodies, and theactivity in vitro was identified by ELISA, flow cytometry and bio-layer interferometry(BLI).ResultsSeven anti-LILRB2monoclonal antibodies with high purity were successfully prepared by hybridoma technique, all of which could recognizeLILRB2 protein with high specificity. Among them, 11C6-8, 43H7-2 and 53A6-11 had superior performance, with the EC_(50)values of 0. 272 3, 0. 431 8 and 0. 344 0 μg/mL, respectively. The chimeric antibodies obtained by humanized modificationexhibited higher binding ability to LILRB2 and effectively blocked the binding of LILRB2 to its ligand, human leukocyteantigen-G(HLA-G), among which the IC_(50)values of 11C6-8 and 53A6-11 were 0. 429 2 and 0. 283 6 μg/mL.ConclusionThe specific monoclonal antibodies against LILRB2 were successfully prepared, and expected to be developed into new anti-tumor immune antibody drugs, which provides a new idea for the development of therapy strategy targeting tumors.
