Nano aptamer for multispecific antibody delivery and its application and construction method
A multi-specific antibody and construction method technology, applied in the field of nano-adapters for multi-specific antibody delivery, can solve the problems of short half-life, high complexity of structure design, and increased complexity, and achieve simple structure design, simple and convenient valency, enhance the effect of antibody efficacy
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Embodiment 1
[0085] Example 1 Construction and Characterization of Nano-Aptamer αFc-NP
[0086] Raw material source and processing method used in the embodiment:
[0087] Goat anti-rat IgG Fc fragment antibody was purchased from Rockland Company, the United States;
[0088] Amino-functionalized polystyrene microspheres (25-300nm), purchased from Shanghai McLean Biochemical Technology Co., Ltd.;
[0089] Sodium Periodate (NaIO 4 ), ready-to-use and ready-to-use, avoid light (purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., China);
[0090] Sodium borohydride, ready-to-use and ready-to-use (purchased from Sarn Chemical Technology (Shanghai) Co., Ltd., China);
[0091] Purpald solution: 10 mg / mL 4-amino-3-hydrazino-5-mercapto-1,2,3-triazole (purchased from Bailingwei Technology Co., Ltd.), dissolved in 1N NaOH. Prepared before use. It reacts with aldehyde groups and becomes purple after the action of sodium periodate (Purpald: 4-amino-3-hydrazino-5-mercapto-1,2,4-triazol...
Embodiment 2
[0119] Example 2, Application of Double Multispecific Nanobodies
[0120] Both the mouse B16-F10 melanoma cell line and the mouse 4T1 orthotopic breast cancer cell line were obtained from the American Standard Biological Collection (ATCC). SPF grade female C57BL / 6 mice and female BALB / C mice, 5-6 weeks old, were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd. The mice were kept in the Experimental Animal Center of South China University of Technology, and the animal experiment procedures followed the relevant provisions of the South China University of Technology's Regulations on the Management of Experimental Animals.
[0121] Rat anti-mouse PD1 (CD279) antibody (αPD1), rat anti-mouse PDL1 (B7-H1) antibody (αPDL1) and rat isotype control (anti-trinitrophenol) antibody (IgG2a): all purchased from the United States Bio X Cell Corporation.
[0122] 1. Binding of bispecific nanobodies to tumor cells and T cells
[0123] 1. In order to simulate the tumor microen...
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