Method for extracting IgM from plasma waste
A waste and plasma technology, applied in the field of protein extraction, can solve the problems that the advantages of IgM cannot be truly reflected, limit the wide application, and low IgM content, and achieve the effects of reducing production costs, improving safety, and reducing IgA content
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Embodiment 1
[0024] Experimental steps:
[0025] (1) Take 0.417kg of FIII precipitate in a 5L blue cap bottle, add 0.05M, pH3.7 acetic acid-sodium acetate buffer solution to make up to 5L, stir and dissolve at 25°C for 4 hours, and filter with a 60LP filter;
[0026] (2) The supernatant is adjusted to pH 4.5 with 0.5mol / L sodium hydroxide solution, and the volume is accurately measured;
[0027] (3) Add n-octanoic acid (1.5% w / v) to the protein solution, stir while adding, stir at 22-24.6°C for 2 hours, continue stirring for another 2 hours and cool down to the final temperature below 5°C with ice-water mixture;
[0028] (4) Take 300ml of protein solution, 10 parts in total, add two parts of diatomaceous earth according to 0%, 0.1%, 0.3%, 0.5%, and 1.0%, respectively, half of which are directly filtered with a wet 50sp filter plate, and the other half Use the balance solution to wet the filter plate and filter, the balance solution is 0.05M acetic acid-sodium acetate buffer pH4.5, 0.1% di...
Embodiment 2
[0036] Experimental steps:
[0037] (1) Take 0.417kg of FIII precipitate in a 5L blue cap bottle, add 0.05M, pH4.0 acetic acid-sodium acetate buffer solution to make up to 5L, stir and dissolve at 25°C for 4 hours, and filter with a 60LP filter;
[0038] (2) The supernatant is adjusted to pH 4.5 with 0.5mol / L sodium hydroxide solution, and the volume is accurately measured;
[0039] (3) Add n-octanoic acid (1.5% w / v) to the protein solution, stir while adding, stir at 25.3-28°C for 2 hours, continue stirring for another 2 hours and cool down to the final temperature below 5°C with ice-water mixture;
[0040] (4) Divide the protein product into two parts equally, add 1.0% diatomaceous earth to one part, stir in the ice-water mixture for 0.5h, and keep stirring the other part in the ice-water mixture for 0.5h;
[0041] (5) Filter the supernatant respectively, concentrate with an ultrafilter with a molecular weight cut-off of 50,000, and the final volume is 97ml.
[0042] The t...
Embodiment 3
[0048] Experimental steps:
[0049] (1) Take four parts of 0.417kg FIII precipitate and place them in 5L blue cap bottles respectively, add 0.05M, pH4.3 acetic acid-sodium acetate buffer solution to make up to 5L, stir and dissolve at 25°C for 4 hours, and filter with a 60LP filter pile;
[0050] (2) supernatant is adjusted pH4.5 with the sodium hydroxide solution of 0.5mol / L;
[0051] (3) According to 1.0%, 1.5%, 2.0%, and 2.5% of the volume, calculate and add n-octanoic acid respectively, stir while adding, control the temperature at 23-28°C and stir for 2h, then continue to stir for 2h while cooling down to The final temperature is below 5°C;
[0052] (4) Filter the supernatant respectively, concentrate with an ultrafilter with a molecular weight cut-off of 50,000, and measure the volume after topping, and detect IgA, IgM, IgG, and total protein content.
[0053] The test results are shown in Table 3 below:
[0054] table 3
[0055]
[0056]
[0057] Experimental ...
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