Preparation Of Genetically Engineered Fusion Protein Vl-Ldp-Ae Of Single Domain Antibody And Lidamycin And Its Anti-Tumor Effect
Keywords
Single Domain Antibodies, Lidamycin, Fusion Proteins, Antibody Drugs, Type Iv Collagenase
Abstract
Type IV collagenase is closely related to tumor invasion, metastasis, and neovascularization. As a tumor-targeting carrier, single-domain antibodies have many advantages that traditional antibodies cannot match. Using type IV collagenase as a target, single-domain antibodies The antibody was used as the carrier, and the potent enediyne anti-tumor antibiotic lidamycin (LDM) was used as the “warhead”. A new method combining DNA recombination and molecular assembly was used to prepare the light chain of the anti-type IV collagenase monoclonal antibody 3G11. The genetically engineered enhanced fusion protein VL-LDP-AE of the variable region single domain antibody VL and LDM. MTT results show that VL-LDP-AE has a strong killing effect on tumor cells cultured in vitro, with an IC50 of HT-1080 cells and KB cells. The values are 8.55¡Á10-12 and 1.70¡Á10-11mol/L respectively. VL-LDP-AE can also inhibit the formation of new blood vessels in the allantoic membrane of chicken embryos and destroy human umbilical vein endothelial cells (HUVEC) to form endothelial tubules. Animals The experiment was conducted to observe the efficacy of transplanted liver cancer 22 (H22) in mice. The tumor was inoculated subcutaneously, and administration began 72 hours later (two intravenous injections on the 3rd and 10th days after inoculation). Comparisons were made with the maximum tolerated dose, VL -The tumor inhibition rate of LDP-AE0.375mg/kg was 89.5%, the tumor inhibition rate of LDM0.05mg/kg group was 69.9%, and the tumor inhibition rate of mitomycin (MMC) 1mg/kg group was 35%.VL – The molecular weight of LDP-AE is 25.4kD, which is much smaller than other forms of antibody drug molecules. As a small, highly efficient antibody drug, VL-LDP-AE has good application prospects in targeted tumor therapy. In addition, its unique preparation method It can also be used as a technical platform for the preparation of new anti-tumor antibody-targeted drugs.
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Original research was done by Miao Qingfang, Shang Boyang, Ouyang Zhigang, Liu Xiaoyun, Zhen Yongsu
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