Monoclonal antibody of chicken infectious laryngotracheitis virus gJ protein and application thereof
A technology of laryngotracheitis virus and monoclonal antibody, which is applied in the field of cell engineering, can solve problems such as difficulty in entering the clinical application stage, high requirements for test conditions and operators, and high test costs
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[0020] The preparation of embodiment 1 monoclonal antibody
[0021] 1. Purification and detection of recombinant protein
[0022] The induced protein rgJ was harvested after 5 hours, centrifuged at 6,000 rpm for 10 minutes, and 10 mL of binding buffer was added. 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 centrifuge at 11,000rpm for 20min 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. The recombinant protein was purified using Bio-RAD's electroelution purification system, and the specific purification steps were as follows:
[0023] (1). Put on gloves and soak the membrane cup in the elution buffer at 60°C for at least 1 hour;
[0024] (2). Plug the filter plate to the ...
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