Monoclonal antibody resisting anaplasma marginale MSP5 protein and application thereof
A technology of monoclonal antibody and marginal anplasma disease, which is applied in the field of cell engineering and can solve problems such as limiting the application of monoclonal antibodies
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[0022] The preparation of embodiment 1 monoclonal antibody
[0023] 1. Purification and detection of recombinant protein
[0024] The induced protein rMSP5 was harvested after 5h, 6000rpm·min -1 Centrifuge for 10min, add 10mL of binding buffer. Resuspend the bacterial liquid, place the sample in an ice bath and ultrasonically lyse (30% power) lyse for 3 seconds with an interval of 15 seconds, and sonicate until the liquid becomes transparent. Then 11000rpm·min -1 Centrifuge for 20 minutes to remove cell debris, and transfer the supernatant to a new tube. Take the supernatant and precipitate separately for SDS-PAGE to determine whether the expressed protein is a soluble protein. Using the His-tag selective tag carried by the pQE30 vector, the protein was purified using Novagen’s instructions. The specific purification steps are as follows:
[0025] 1) Mix 3mL sterile deionized water with the resin, and suck out the supernatant after the resin sinks completely. Wash twice....
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