Orf virus (ORFV) protein ORFV059 monoclonal antibody hybridoma G3 and monoclonal antibody
A technology of ORFV059 and aphthous ulcer virus, which is applied in the field of cell engineering, can solve the problems of ineffective research on disease control and lack of specific antibodies, and achieve the effects of reduced application cost, high specificity and sensitivity
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[0021] Example 1 Preparation and identification of the monoclonal antibody of the present invention
[0022] 1. Preparation of monoclonal antibody G3
[0023] 1) Preparation of recombinant ORFV059 antigen
[0024] Using the orf virus genome as a template, the full-length ORFV059 gene was obtained by PCR amplification, and the prokaryotic recombinant expression vector pET28a(+) / ORFV059 was constructed, which was expressed in Escherichia coli Escherichia coli Expressed in BL21, and purified by Ni Sepharose affinity chromatography purification column, the recombinant ORFV059 protein with a purity of more than 90% was obtained.
[0025] 2) Immunization of mice
[0026] 6-8 week old female BALB / c mice were immunized with purified recombinant antigen.
[0027] The first immunization: After mixing the recombinant antigen with an equal volume of Bentonite adjuvant, inject BALB / c mice intraperitoneally at 50 μg / 500 μL each;
[0028] The second immunization: After 2 weeks, take th...
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