Application of a monoclonal antibody in the treatment of neurodegenerative diseases
A monoclonal antibody and neurodegenerative technology, applied in the direction of nervous system diseases, antibodies, nervous system cells, etc., can solve the problems of inability to alleviate pathological damage of neuroinflammation and complicated pathogenesis
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Embodiment 1
[0125] Example 1 Affinity determination of recombinant human anti-tumor necrosis factor-α monoclonal antibody and soluble TNF-α
[0126] 1. Experimental materials
[0127] Experimental drug: recombinant human anti-tumor necrosis factor-α monoclonal antibody injection, the sequence of which is shown in SEQ ID NO.: 1, stored in a refrigerator at 4°C.
[0128] Recombinant human TNF-α: purchased from Shanghai Nearshore Protein Technology Co., Ltd., item number: C008, batch number: 0327018.
[0129] Amine Coupling Kit (Lot No. 2057704), HBS-EP+buffer (10×) (Lot No. BCBJ0266V), Human Antibody Capture Kit (Lot No. 10137214): all purchased from GE Healthcare.
[0130] Biacore T200, CM5 sensor chip (Serial No. 10148134): purchased from GE Healthcare.
[0131] 2. Experimental method
[0132] 2.1 Solution preparation
[0133] Mobile phase buffer solution: HBS-EP + Buffer solution (10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% P20, pH 7.4).
[0134] Sample dilution solution: HBS-EP + ...
Embodiment 2
[0145] Example 2 Recombinant human anti-tumor necrosis factor-alpha monoclonal antibody binding specificity
[0146] 1. Experimental materials
[0147] Test product: recombinant human anti-tumor necrosis factor-α monoclonal antibody injection, the sequence of which is shown in SEQ ID NO.: 1, stored in a refrigerator at 4°C.
[0148] Recombinant human TNF-β: purchased from Sino Biological Inc., product number: 10270-HNAE, lot number: LC05JL0410; recombinant monkey TNF-α: purchased from Sino Biological Inc., product number: 90018-CNAE, lot number: LC06DE1213; recombinant dog TNF-α: purchased from Sino Biological Inc., product number: 70003-DNAE, lot number: LC05JU2002; recombinant mouse TNF-α: purchased from Shanghai PrimeGene Bio-Tech Co., Ltd. company, product number: 123-01, lot number : 810588; recombinant rat TNF-α: purchased from Shanghai PrimeGene Bio-Tech Co., Ltd. company, item number: 143-01, batch number: Y14301011; goat anti-human IgG-HRP absorbed by monkey serum wi...
Embodiment 3
[0156] Example 3 Inhibitory Effect of Recombinant Human Anti-TNF-α Monoclonal Antibody on the Binding of TNF-α to Its Receptor
[0157] 1. Experimental materials
[0158] Test product: recombinant human anti-tumor necrosis factor-α monoclonal antibody injection, the sequence of which is shown in SEQ ID NO.: 1, stored in a refrigerator at 4°C.
[0159] Recombinant human TNF-α was purchased from Shanghai Jinan Protein Technology Co., Ltd., product number: C008, batch number: 0327018; recombinant TNF RI (p55) and TNF RII (p75) were purchased from R&D Company, product number 372-R1-050 and 726, respectively -R2-050, the batch numbers are APJ0712071 and BVD0312111 respectively; goat anti-human IgG-Fc was purchased from Kirkegaard & Perry Laboratories, Inc., product number: 01-10-20, batch number: 110078; BSA was purchased from Roche Company, batch number: 10735078001.
[0160] 2. Experimental steps
[0161] (1) Dilute 1 mg / ml goat anti-human IgG-Fc to 10 μg / ml with PBS, add 50 μl...
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