1.Dilution of Antigen: Dilute the antigen with physiological saline to twice the final concentration (prepared according to an antigen dose of 1 ml per injection). Recommended antigen doses:
•(1) For subunit protein antigens with weak immunogenicity, 800 μg per injection.
•(2) For inactivated whole virus or whole bacteria and virus-like particle antigens with strong immunogenicity, 400 μg per injection.
•(3) For small molecule antigens conjugated to carrier proteins, 500 μg per injection. (Actual dosage should be based on preliminary data and company experience.)
2.Mixing the Adjuvant: Thoroughly mix the adjuvant (it is recommended to use a syringe to mix repeatedly), and under sterile conditions, take the required amount (1 ml per injection) and mix quickly with the antigen in a 1:1 volume ratio. (Slight precipitation of the adjuvant is normal; ensure thorough mixing with the antigen and inject promptly.)
3.Injection Method: Inject into the muscle of the leg, administering 2 ml per goat. (The injection method can follow the company's routine operations.)
•(1) Slight precipitation of the adjuvant after mixing with the antigen is normal. Ensure thorough mixing before drawing into the syringe and inject promptly.
•(2) Alternatively, subcutaneous or intradermal injection can be chosen based on experimental habits.
4.Booster Immunization: On the 10th-14th day, administer a booster injection with the same dose and method (the timing of the second injection should be determined based on the type of antigen). Note: Always prepare and use the adjuvant and antigen freshly mixed, injecting at the same site as the initial immunization. (Follow the company’s actual immunization schedule for booster immunizations.)
5.Antibody Detection: IgG can be detected on the 14th day, and a small blood sample can be collected on the 21st day for ELISA testing. (The above is a predicted schedule; actual data conclusions should follow the experimental results.) The antibody titer can reach its peak. Then, whole blood can be collected, or antigen challenge immunization and spleen cell fusion can be performed following standard methods.
6.Additional Booster if Needed: If the titer on the 21st day is below expectations, an additional booster shot with the same dose and method can be given around the 28th day. Titer testing can be performed on the 28th-35th days, generally reaching peak antibody titer. Subsequently, whole blood collection or antigen challenge immunization and spleen cell fusion can proceed following standard methods.
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