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An antibody and antigen technology, applied in the direction of antibodies, applications, anti-tumor drugs, etc., can solve the problem of low anti-FGL1 antibodies, achieve high binding activity, good stability, and enhance the effect of anti-tumor effects
Active Publication Date: 2022-07-05
SHANDONG BIOANTY BIOLOGICAL TECH CO LTD
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Problems solved by technology
However, in the prior art, there are few reports about anti-FGL1 antibodies
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Embodiment 1
[0041] Example 1. Generation of anti-FGL1 monoclonal antibodies
[0042] 1.1 Immunization
[0043] The fully human antibody transgenic mice BoAn-hMab (Shandong Boan Biotechnology Co., Ltd.) were immunized with FGL1 (Bepsis, FG1-H52Hy) and mGM-CSF (Yiqiao Shenzhou, 51048-M07H) and Freund's adjuvant after emulsification. ). A total of 11 mice were immunized with Freund's complete adjuvant for the first immunization, and Freund's incomplete adjuvant for the second to fifth immunization, all of which were injected subcutaneously. 11 mice were boosted by intraperitoneal injection of the same dose of antigen and adjuvant with the second to fifth immunization, and sacrificed 3 days later, and the spleen was taken out for phage library establishment.
[0044] 1.2 Establishment of phage library
[0045] Take the spleen cells of the immunized mice, extract RNA, and then reverse-transcribe to obtain cDNA. The establishment of phage library is carried out according to the method descri...
Embodiment 2
[0073]Example 2. PK activity of CA352 antibody in mice
[0074] Take 3 BALB / c mice, after intravenous injection of 5mg / kg CA352 antibody, before administration (0 h) and 5min, 30min, 1h, 3h, 6h, 24h, 48h, 96h, 168h, 240h after administration , 336h, 384h, and 432h, the blood was collected from the mouse orbit, the blood sample was placed in a collection tube, and allowed to coagulate naturally in an ice box. After the blood sample was taken out, 8h was built in a centrifuge, and centrifuged at 1000-3000 g for 10min to separate the serum. In the sample storage tube, Elisa was used to detect the metabolism of CA352 antibody in BALB / c mice. The results are shown in image 3 and Table 7. from image 3 It can be seen from the PK data of CA352 in BALB / c mice in Table 7 that the half-life of CA352 in vivo is 118 hours, which is relatively stable.
[0075] Table 7
[0076]
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Abstract
The present invention relates to a FGL1 antibody or an antigen-binding fragment thereof. The FGL1 antibody or its antigen-binding fragment has high binding activity to FGL1 protein, and can realize the blocking of the binding of FGL1 protein and LAG3 protein, and the combination of FGL1 protein and expression. Blockade of cellular binding of LAG3 protein. The present invention also relates to a nucleic acid encoding the FGL1 antibody or an antigen-binding fragment thereof; a cell containing the nucleic acid; a composition containing the FGL1 antibody or an antigen-binding fragment thereof, the nucleic acid, the cell; containing the FGL1 Kits for antibodies or antigen-binding fragments thereof, nucleic acids, and compositions; and the FGL1 antibody or antigen-binding fragments thereof, nucleic acids, and compositions in the preparation of reagents for detecting, diagnosing tumors, or treating tumors. application in medicine.
Description
technical field [0001] The present invention relates to the field of biotechnology, in particular to an anti-FGL1 antibody and use thereof. Background technique [0002] LAG-3, lymphocyte activation gene-3 (Lymphocyte activation gene-3) belongs to the immunoglobulin superfamily and is a type I transmembrane protein composed of 498 amino acids, which can be selectively expressed in activated T lymphocytes , NK cells and dendritic cells. The regulatory function of LAG-3 protein on T cells is similar to that of PD-1, mainly acting as a receptor for delivering inhibitory signals. [0003] At present, most scientists believe that MHC-II is the main ligand used by cancer cells to bind LAG-3 molecules and thereby reduce the activity of T cells. Therefore, blocking the binding of LAG3 to MHC-II will help activate the anti-cancer ability of T cells. However, , several monoclonal antibodies that do not block the binding of LAG-3 to MHC-II have shown the ability to activate T cells, ...
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