Sequences of variable regions of anti-GFAP (glial fibrillary acidic protein) monoclonal antibody and method for preparing same
A monoclonal antibody and variable region technology, applied in the field of biomedicine, can solve problems such as the reduction of GFAP expression level
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[0028] Example 1. Preparation and Identification of Anti-GFAP Monoclonal Antibody
[0029] 1. Preparation of hybridoma cells
[0030] (1) Animal immunization: Simultaneously immunize 3 healthy female BALB / c mice aged 6-8 weeks with GFAP protein antigen. After the 3 times of immunization, blood was collected once a week, and the OD value detected by ELISA at 1:4000 dilution was selected to be greater than The serum of 1.0 and BALB / c mice with positive serum WB test were used for fusion, and 3 days before the fusion, the antigen without adjuvant was injected intraperitoneally, and the injection dose was 50ug / mouse.
[0031] (2) Collect B lymphocytes: 3 days after the booster immunization, take blood from the mice, centrifuge the blood, and keep the serum as a positive control; then take the spleen of the mouse under sterile conditions, and put the spleen in 10ml pre-warmed incomplete In the culture medium, peel off the surrounding connective tissue, place it in a 100-mesh stain...
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