Preparation method of dry powder of recombinant human interferon-alpha-2b-BCG

Abstract

The invention discloses a preparation method of dry powder of recombinant human interferon-alpha-2b-BCG and belongs to the technical field of biological engineering. The preparation method comprises the following steps of: inoculating a recombinant human interferon-alpha-2b-BCG strain into a Middlebrook7H9 liquid culture medium to obtain a first-level seed solution, wherein 1 milliliter of the Middlebrook7H9 liquid culture medium contains 30 microliters of kanamycin; then transferring in the liquid culture medium to obtain a second-level seed solution with the A value of concentration standard OD600 being more than 2; carrying out large-scale submerged fermentation culture by using a biological reaction generator, connecting a fermentation tank to a tangential flow hollow filtering column in situ, washing thalli in a dispersing way for fermentation liquor, and then adding the same volume of mixed liquor which contains 20% of sucrose, 1%-2% of gelatin, 2% of potassium chloride and 1%-4% of sodium glutamate for protection solution displacement; slowly precooling to reduce temperature, and carrying out vacuum freeze-drying to obtain the dry powder. The recombinant human interferon-alpha-2b-BCG obtained in such a way is high in bacterin activity rate and reduces the possibility of pollution in a production process.

Description

technical field [0001] The invention relates to bioengineering technology, in particular to a dry powder preparation method of recombinant human interferon-α-2b-BCG. Background technique [0002] Recombinant human interferon-α-2b-BCG (hIFN-α-2b-BCG) is a biologic agent currently used for immunoperfusion therapy of bladder tumors. [0003] In the past, the freeze-drying preparation of BCG seedlings used the method of static cultivation of Sutong potatoes, extrusion of the bacterial membrane to remove water, and freeze-drying after grinding in steel cylinders. , Bile Potato Medium or Liquid Sutong Medium for static culture for 2 to 3 weeks, culture and passaging activation. Then, carry out production culture: pass another generation on the Sutong potato medium or directly pick the well-grown bacterium film, inoculate on the surface of Sutong comprehensive medium or other approved medium, in the culture bottle, 37 ~39°C constant temperature, static culture for 7~14 days. Th...

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Problems solved by technology

[0006] The present invention aims to solve the problems in the prior art that the production process of recombinant interferon-α-2b-BCG freeze-dried powder is complex in operation, slow in growth, low in viable bacteria rate, prone to pollution, and poor in process controllability, etc., and provides a Dry powder preparation method of recombinant human interferon-α-2b-BCG

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