Diagnostic marker for systemic lupus erythematosus
A lupus erythematosus, systematic technology, applied in the direction of material inspection products, measuring devices, instruments, etc., can solve problems such as easy missed diagnosis
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Embodiment 1
[0033] Using the PNGase F enzyme kit (IBL, Germany), according to the kit instructions provided by the manufacturer, the N-glycan chains of serum antibodies were extracted, and the N-glycan chains of serum antibodies were analyzed by primary and multi-stage mass spectrometry. The process is as follows:
[0034] (1) Collect serum samples from SLE patients and centrifuge at 2000r / min for 10min.
[0035] (2) Take the supernatant, dilute it at the ratio of 1mL serum to 3mL PBS buffer, add saturated ammonium sulfate buffer dropwise, let stand for 3h, and then centrifuge at 6000r / min.
[0036] (3) The precipitate obtained in step (2) was further desalted with sephadex G25, and then passed through a proteinG column to further purify the antibody, and freeze-dried to obtain serum antibody, which was stored at -20°C for future use.
[0037] (4) Put 500 micrograms of serum antibody into a centrifuge tube, add 5 microliters of 10× denaturant and 45 microliters of ultrapure water, mix wel...
Embodiment 2
[0058] Construct the SLE mouse model:
[0059] BALB / c mice aged 6-8 weeks were purchased and raised in an SPF animal room. After the environment was stabilized, they were randomly divided into two groups, with 5 mice in each group. The mice in the experimental group (ie, the SLE model group) were intraperitoneally injected with 0.5 mL of pristane (purchased from sigma), while the mice in the control group were injected with 0.5 mL of PBS. Blood was collected once a week before and after the injection, and ELISA and other methods were used to detect the contents of anti-dsDNA, anti-sm RNP and anti-ribosomal PO antibodies, and finally confirmed the success of the establishment of the SLE mouse model.
[0060] Treatment: Feed the mice for about 6 months, kill them, and take the serum for future use. According to the method in Example 1, N sugar chains were extracted, and after primary and multi-stage mass spectrometry detection, the primary mass spectrogram was as follows Figu...
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