Mitochondria creatinekinase antibody
A creatine kinase and mitochondrial technology, applied in the field of clinical examination, can solve the problems of lack of anti-umCK antibody and difficulty in distinguishing
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Embodiment 1
[0082] Example 1: Production of hybrid cell tumors producing monoclonal antibodies (MoAb)
[0083] (1) Modulation of immunogen (antigen)
[0084] The present invention uses human umCK as an antigen (Lot No. ETH010122 manufactured by the Institute of Cell Biology).
[0085] (2) Place the immunized 5-8 week-old inbred BALB / c female mice in the animal room (23±1°C, temperature 70%), feed them with standard granules, and give them water arbitrarily.
[0086] (3) The refined product prepared in the above (1) with human umCK as the antigen was prepared with PBS to a concentration of 100μg / 0.5mL and an equal amount (0.5mL) of Freund's complete adjuvant (frend's complete adjubant: manufactured by Difco) was mixed and emulsified. Inject 200 μL of this emulsified antigen into the abdominal cavity of 5-week-old female BALB / C mice, and then inject 100 μg / mL of the above-mentioned antigen prepared with Ribi adjuvant into each mouse every 2 weeks. Inject 20 μg, four times in total. Then...
experiment example 1
[0101] Experimental example 1: Regarding the specificity of enzyme repression of culture supernatant
[0102] With respect to the culture supernatant of the hybrid cell tumor-producing cells in the 7 acupoints screened above, inhibition of each enzyme activity was investigated using human umCK, human smCK, human CK-MM, human CK-BB, or human CK-MB, and the effect on the activity of each enzyme was confirmed. Specificity of human CK isozymes. For verification, a MoAb of mCKI-578 (JP-A 2002-270 publication) produced by a hybrid cell tumor commissioned under the commission number FERM BP-7133 was also investigated.
[0103] The result is as figure 1 shown. The supernatants of pure lines UI-178, UI-281, UI-956, UI-1111, UI-1125, UI-1881 and UI-1299 can inhibit more than 82% of umCK enzyme activity, but for smCK, CK- The enzymatic activities of MM, CK-BB, and CK-MB were not suppressed at all. Especially the supernatant of UI-1881 could repress 94% of umCK enzyme activity. On th...
experiment example 2
[0104] Experimental Example 2: Establishment of a MoAb Hybrid Cell Tumor Line (Selection Establishment)
[0105] The above-mentioned hybrid cell tumors in the 7 cavities obtained by screening were screened by the limiting dilution method, and one pure line was selected that could stably produce hybrid cell tumors showing inhibition of mCK enzyme activity. Using this cell as the establishment strain, we entrusted the processing to the Institute of Bioengineering and Industrial Technology, Institute of Industrial Technology, Ministry of International Trade and Industry, under the consignment number FERM P-18760.
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