Method for measuring content of rhCNB (Recombinant Human Calcineurin B)
A content and external standard method, applied in the biological field, can solve the problems of inaccurate results and low anti-interference ability, and achieve the effect of good specificity and high accuracy
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Embodiment 1
[0060] Take an appropriate amount of this product (6.0mg of pure recombinant human calmodulin phosphatase B subunit), weigh it accurately, put it in a 10ml measuring bottle, add 1mol / L reduced DTT solution (dithiothreitol), reduced gluten Cycloside, β-mercaptoethanol, sodium borohydride or sodium bisulfite or a mixture of two or more 80μl, sodium hydroxide test solution 40μl, add about 8ml of water, react at 80℃~100℃ for 15~60min , take it out and let it cool to room temperature (15-35°C), add water to dilute to the scale to make a test solution with a protein content of about 0.6 mg / ml, accurately measure 100 μl, inject it into a liquid chromatograph, and record the chromatogram.
[0061] Another bottle of calibrated recombinant human calmodulin phosphatase B subunit working reference substance was taken out, and an appropriate amount (6.0 mg of pure recombinant human calmodulin phosphatase B subunit) was accurately weighed, and determined in the same way, according to the ext...
Embodiment 2
[0064] Test sample: protein monomer solution containing TRIS, EGTA and metal ions 7.06mg / ml
[0065] Mainly for the defects of the Folin phenol method (the sample must not contain Tris, reduced DTT, mercaptoethanol, EDTA, etc.)
[0066] Also for the defects of the biuret method (samples must not contain Tris, EDTA, etc.);
[0067] Measure according to the detection method of embodiment 1:
[0068] figure 1 The spectrogram data are shown in Table 1:
[0069] Table 1 figure 1 The spectrogram data of
[0070]
Embodiment 3
[0072] Sample to be tested: rhCNB protein monomer solution 12.13mg / ml containing 10mmol citrate buffer
[0073] For the defects of the C18 column high-performance liquid phase method for measuring rhGM in the three appendices of the 2010 edition of "Chinese Pharmacopoeia", (proteins with the same hydrophobicity cannot be separated, and in this method, they are completely separated due to different molecular weights) according to the detection of Example 1 Method determination:
[0074] figure 2 The spectrogram data of are shown in Table 2:
[0075] Table 2 figure 2 The spectrogram data of
[0076]
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