Fed batch culture method for protein secreting cells

Details Fed batch culture method for protein secreting cells Fed batch culture method for protein secreting cells

1994-01-03

1995-08-22

Lilling, Herbert J.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Using tissue cell culture to make a protein or polypeptide

43524025, 4352402, C12N 508, C12P 2100

Patent

active

054439680

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

1. Field of the Invention
This invention relates to a process for the production of a useful protein by making cells secrete the useful protein. More particularly, this invention relates to a process which comprises culturing a human embryonal kidney-derived 293 cell transformed so as to express a useful protein to obtain a culture fluid containing the desired useful protein in a high concentration, and recovering the useful protein from the culture fluid.
2. Description of Related Art
Production of useful proteins, particularly physiologically active proteins by culturing animal cells, is widely carried out. Particularly, processes of producing useful proteins by culturing animal cells transformed so as to secrete the useful proteins are indispensable techniques for production of certain useful proteins.
Animal cells are superior to bacteria and yeasts in the ability of post-translational modification. However, when animal cells are used, the cost of the medium and purification are relatively high due to a low product concentration in culture fluid. Therefore, it is important for industrial production to raise the concentration of useful proteins secreted by animal cells. In a simple batch culture method, proliferation of the cells stops at some point of the culture period mainly due to the exhaustion of nutrients, and secretion of the protein also stops. In order to improve this simple batch culture method, a so-called fed-batch culture method is carried out in which exhausted nutrients, for example sugars, amino acids or a fresh medium, are added continuously or intermittently during the culture period.
In this specification, a "simple batch culture method" means a method which comprises inoculating cells into a medium in a culture vessel to start culture, and carrying out the culture without substantial addition of part or all of the nutrients or a new medium until completion of the culture, and a "fed-batch culture method" means a method comprising inoculating cells into a medium in a culture vessel to start culture, and carrying out the culture while adding part or all of the nutrients or a fresh medium continuously or intermittently into the culture vessel, without substantially taking out the culture fluid from the culture vessel.
It is known that in a fed-batch culture method, the culture period is prolonged and higher cell density and product (protein) concentration can be obtained compared to a simple batch culture method (J. B. Griffith, Animal Cell Culture and Production of Biologicals, pp. 401-410, Klumer Academic Publishers, R. Sasaki and K. Ikura (eds.), (1991), S. Reuveny et al., J. Immunol. Methods, 86, 53 (1986), S. Reuveny et al., J. Immunol. Methods, 86, 61 (1986)). In the above literature of J. B. Griffith, a comparison shown in the following Table 1 is given as a typical example.


TABLE 1 ______________________________________ Number of Product Culture cells yield Per liter Length type (millions) (mg/week) (mg/month) (days) ______________________________________ Batch 3 100 200 7 Fed 6 200 500 14 batch ______________________________________
The above S. Reuveny et al. compares a simple batch culture with a fed-batch culture in the latter of the above literatures. This comparative experiment has been conducted under the following conditions: (1) The simple batch culture comprised inoculating hybridoma cells at a density of 3.times.10.sup.5 cells/ml in 100 ml of a medium and carrying out culture for 8 days, and (2) the fed-batch culture comprised inoculating hybridoma cells at a density of 3.times.10.sup.5 cells/ml in 60 ml of a medium, and then carrying out the culture for 8 days while adding 6 ml of a fresh medium once a day after the density had become about 1.times.10.sup.6 cells/ml 2 days after inoculation. The results show that the antibody productivities in both methods are 15 mg/l/day and 27 mg/l/day, respectively. Thus, S. Reuveny et al. teaches an about 1.8-fold increase of the productivity (antibody) in the fed-batch cult

REFERENCES:
patent: 5219752 (1993-06-01), Takazawa
patent: 5328844 (1994-07-01), Moore
Journal of Immunological Methods, 86 (1986) pp. 61-69, "Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors" S. Reuveny et al.

Affiliated with

Also associated with

Rating

Fed batch culture method for protein secreting cells does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Fed batch culture method for protein secreting cells, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Fed batch culture method for protein secreting cells will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2141239