Anti-aβ31-35 antibody for treating and preventing Alzheimer's disease and preparation method thereof
An Alzheimer's disease and antibody technology, applied in the field of anti-Aβ31-35 antibody and its preparation for the treatment and prevention of Alzheimer's disease, achieving the effect of small side effects and strong specificity
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Embodiment 1
[0015] Preparation steps of anti-Aβ31-35 antibody for treating and preventing Alzheimer's disease:
[0016] (1) Antigen preparation
[0017] The Aβ31-35 fragment was synthesized by organic solid-phase synthesis method, the solid phase carrier was HMP resin, the amino acid protected by Fmoc (9-fluorenylmethyloxycarbony) was used to synthesize the amino acid from the carboxyl end of the polypeptide to the amino end, and the peptide resin was cleaved by TFA method. Crude Aβ31-35 fragments were obtained after pressure distillation and ether extraction. It was then purified by high-performance liquid chromatography (HPLC), and the purified sample was analyzed by HPLC. The purity was 95%, and the purified Aβ31-35 fragment was freeze-dried.
[0018] Take the purified Aβ31-35 fragment and couple with hemocyanin. The coupling method is to use the carbodiimide method [1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide, EDC] for condensation reaction to obtain Aβ31-35 Fragment-hemocyanin c...
Embodiment 2
[0024] We observed the protective effect of anti-Aβ31-35 antibody on brain function or cell damage caused by intracerebroventricular injection of Aβ or addition of Aβ to neuronal cell culture. The main results are as follows:
[0025] (1) Morris water maze experiment figure 2 The average latency (A) and typical swimming trajectories (B) of rats in each group to find the underwater platform are shown. Compared with normal rats, intracerebroventricular injection of 5 nmol Aβ1-42 can significantly prolong the platform-seeking latency; after treatment with different concentrations of anti-Aβ31-35 antibody, the latency and swimming distance are shortened, especially 0.5 and 5 nmol anti-Aβ31-35 antibody pre-treatment. The treatment group was significantly smaller than the Aβ1-42 group on each training day (P<0.01), indicating that the behavioral damage caused by Aβ can be reversed by the anti-Aβ31-35 antibody.
[0026] (2) In vivo hippocampal LTP pre-experiment such as image 3 ...
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