A kind of anti-malignant melanoma vaccine composition and its application
A technology of malignant melanoma and vaccine composition, applied in the field of biopharmaceuticals, can solve the problem of increasing the specific anti-tumor immune response of malignant melanoma vaccine, unable to effectively induce or activate the patient's immune response, malignant melanoma specificity and immune response. Weak response and other problems, to achieve the effect of strong specific killing function, good treatment effect and convenient use
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Embodiment 1
[0040] 1. Preparation of malignant melanoma antigen
[0041] (1) Preparation of malignant melanoma attenuated cells
[0042] M3 malignant melanoma cell line (H-2 d )(2xl0 6 / mL) into a 6-well plate, and then 60 Co irradiation 30Gy spare.
[0043] (2) Preparation of DNP-modified malignant melanoma attenuated cells
[0044] M3 malignant melanoma cell line (H-2 d )(2xl0 6 / mL) into a 6-well plate, and then 100L of DNP solution to a final concentration of 0.2% (5mmol / L, pH7.2), incubated at 37°C for 10min, and then 60 Co irradiation 30Gy spare.
[0045] 2. Preparation of antigen mixture containing tuberculosis antigen, malignant melanoma antigen and rod-shaped bodies
[0046] Take the malignant melanoma antigen according to the amount of 80%-100% (volume), take the tuberculosis antigen according to the amount of 0%-20% (volume), and take the rod-shaped bodies according to the amount of 0%-20% (volume) and mix them evenly Backup.
[0047] 3. Preparation and activation of DC...
Embodiment 2
[0088] 1. Preparation of malignant melanoma antigen
[0089] (1) Preparation of malignant melanoma attenuated cells
[0090] The logarithmic growth of malignant melanoma cell line B16 (H-2 b )(1xl0 7 / mL) into a 6-well plate, and then 60 Co irradiation 30Gy spare.
[0091] (2) Preparation of DNP-modified malignant melanoma attenuated cells
[0092] The logarithmic growth of malignant melanoma cell line B16 (H-2 b )(1xl0 7 / mL) into a 6-well plate, and then 100 μL of DNP solution to a final concentration of 0.2% (5mmol / L, pH7.2), incubated at 37°C for 10min, and then 60 Co irradiation 30Gy standby, dose rate 1.88Gy / min.
[0093] 2. Preparation of antigen mixture containing tuberculosis antigen, malignant melanoma antigen and rod-shaped bodies
[0094] Take malignant melanoma antigen B16 attenuated cells according to the amount of 80%-98% (volume), take tuberculosis antigen BCG according to the amount of 2%-10% (volume), and take rod-shaped small Mix well and set aside. ...
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