Application of crmp2 and anti-crmp2 antibody
An antibody and reagent technology, applied in the application field of CRMP2 and anti-CRMP2 antibodies, can solve problems such as unclear mechanism
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Embodiment 1
[0043] The numbers of the 3 patients in this example are: P1, P2, and P3, respectively.
[0044] P1 was the first case, and CRMP2 was identified by its cerebrospinal fluid and blood samples; P2 and P3 were positive cases of anti-CRMP2 antibody detected by retrospective analysis of CBA method.
[0045] P1, P2, and P3 all had symptoms of dizziness or headache, visual impairment, and unsteady walking; white matter lesions; immunotherapy was effective; blood and / or cerebrospinal fluid in rat brain slices were positive for TBA; anti-CRMP2 antibody was positive for other AEs Antibody tests were negative.
[0046] P1 and P3 had language barriers, bilateral finger-to-nose tests were inaccurate; P2 had chest and abdomen pain, and spinal cord lesions.
[0047] P1 cerebrospinal fluid was positive for Mycoplasma pneumoniae; P3 cerebrospinal fluid was positive for human herpesvirus 6 and Klebsiella pneumoniae; blood was positive for influenza B virus IgM.
[0048] The specific clinical d...
Embodiment 2
[0056] Immunohistochemical experiments based on rat brain slices are routine examinations for autoimmune encephalitis, including immunoenzyme chemistry and immunofluorescence (also known as TBA, tissue based assay).
[0057] Implementation process: perfuse the heart of an adult rat with PBS (phosphate buffered saline) after anesthesia to remove circulating blood, take the brain (including the hippocampus) for sagittal frozen section, take the cerebellum for longitudinal section, fix it in cold acetone for 10 minutes, and let it dry Dry and set aside. The slices were washed 3 times with PBST solution (PBS containing 0.05% Tween-20), 3 minutes each time.
[0058] Next, carry out immunoenzyme chemistry method and immunofluorescence method respectively to detect: (1) Carry out immunoenzyme chemistry method: treat with 3% hydrogen peroxide again for 10 minutes, wash 3 times with PBST and carry out next step blocking, use 10% goat serum / PBST solution at room temperature for blocki...
Embodiment 3
[0064] Identification of antigens targeted by autoantibodies in cerebrospinal fluid of P1 patients using immunoprecipitation combined with protein profiling.
[0065] Implementation process: take fresh brain tissue of about 2-month-old rats, add 1 ml protein lysate (20mmol / L Tris-HCl, pH 7.4, 150mmol / L NaCl, 2mmol / L EDTA, 1% TritonX-100, protease inhibitor ), homogenized for 1 minute with a glass homogenizer, and lysed on ice for 30 minutes. Centrifuge at 12,000 rpm at 4°C for 15 minutes, and take the supernatant as rat brain protein lysate. Mix 500 microliters of protein lysate with 500 microliters of cerebrospinal fluid of P1 patient, and vortex vertically at 4°C overnight. Add 50 μl of pre-washed protein A-labeled agarose, and vortex vertically at 4°C for 2 hours. Centrifuge at 4°C, 1000rpm, resuspend, rinse with protein lysate 3 times, then add 1xSDS-PAGE sample buffer, boil for 5 minutes to elute the protein complex. The prepared samples were separated by SDS-PAGE, fol...
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