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Cell suspension culture tank

A cell culture and cell reaction technology, applied in the field of cell culture tanks, can solve problems such as expensive and achieve the effect of extending the stroke

Inactive Publication Date: 2007-12-12
HANGZHOU AMPROTEIN BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, optimizing a control tower or probe is expensive

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0045] Embodiment 1: the research of 150ml wide body flat conical bottom cell reactor

[0046] The fed-batch culture of CHO-producing cell lines in small shake flasks is an important way to screen solid and high-yield producing cell lines. In this study, the results (Table 1) using a 150ml wide-bodied flat conical bottom cell reactor (Figure 2) and a wide-bodied conical bottom shake flask (Corning Company #431123) showed that the 150ml shake flask is suitable for small-scale Fed-batch study, the inoculation volume is small (only 1 / 20 of the working volume). The expression titer can be compared with a 2L reactor. Inverted flat conical shake flasks obviously have cell sedimentation, especially those cell clusters are more likely to settle, mainly because the mixing of the pointed bottom is not good. For this reason, we gave up using the inverted conical bottom and adopted Inverted flat cone bottom.

[0047] Table 1: The culture tank is placed on the shaker platform with a rot...

Embodiment 2

[0050] Embodiment 2: Research on conical bottom tanks with different cone angles

[0051] Plastic centrifuge tubes with different cone angles were used on a shaker to study the effect of cone angle on shaking, medium mixing and inoculum volume (Table 2). If the cone angle is not too wide or too narrow, shaking cannot cause the culture volume to spread to the tank wall to form a wider thin layer to expand the surface of the medium. For example, if the cone angle is too wide, such as when it is greater than 70 degrees, it has no obvious difference from the flat-bottomed tank. If the cone angle is too small, such as when it is less than 30 degrees, it has no obvious difference from the flat-bottomed tank. The results of Table 3 It was shown that a cone angle of 30 degrees was the minimum angle for effective shaking and mixing. According to calculations, tanks with cone angles greater than 70 degrees did not significantly reduce the inoculum volume compared with conventional round...

Embodiment 3

[0055] Embodiment 3: the research of 3 liters of wide-bodied flat conical bottom shake flasks

[0056] The 3-liter wide-body flat-conical bottom cell reactor is equipped with an oxygen tube device or air injection (Fig. 1a, b), and the medium in the working volume has a large surface area, which is conducive to the absorption of oxygen by the medium (per cubic centimeter of medium The surface area of ​​the reactor is greater than 0.143cm2), and the wide-body flat-conical bottom cell reactor is equipped with an air injection device at the bottom to extend the escape journey of the air (containing 21% oxygen) (figure a, b). The oxygen tube device at the bottom is used as a low flow meter to control the low flow rate of pure oxygen. The shaker can make the culture volume spread to the tank wall, and it is easy to form a thin culture layer to extend the culture surface area and improve aeration.

[0057] CHO cell lines were used to express TNFR1-Fc-IL-1ra, IL-18bp-Fc-IL-1ra, VEGFR...

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Abstract

The invention discloses a cell suspension culture tank and its usage. Compared with current flat base biological tank, it is characterized by wide- bodied culture tank, converse flat conical bottom, larger culture medium area, benefit for oxygen absorption and carbon dioxide discharge, reduced cell inoculation volume, much lower level of air injection, prolonged air bubble course, regular agitation that makes culture medium contacts with tank wall, and generates small cutting force and mechanical force, and benefit for surface area enlargement, air aeration amount and improved mixing. The disposal plastic or glass shake flask suspension culture system using no complicate control tower and relevant probe, is developed by using design in this invention. The invention aslo relates to large- size industrial biological reactor which is drove by stirring paddle.

Description

field of invention [0001] The invention relates to a cell culture tank, which has an inverted flat cone bottom or an inverted frustum bottom, which can reduce the cell seeding volume, better mix to avoid mechanical shear force, and have better air flow . Background of the invention [0002] High-density suspension culture of animal cells is a necessary method for the production of protein drugs or vaccine products. Small-volume cell culture tanks are used for laboratory production, and large-volume cell culture tanks are used for industrial production. [0003] The shapes of current cell culture tanks and shake flasks are all typical cylindrical shapes. The height:diameter ratio of the working volume of such a tank or bottle is greater than 1:1. The medium surface area in the tank is small, which is not conducive to the effective absorption of oxygen by the culture. Air or oxygen sparging is commonly used to solve this problem. However, excessive air sparging often prod...

Claims

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Application Information

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IPC IPC(8): C12M3/02
Inventor 惠觅宙李会成冷国庆
Owner HANGZHOU AMPROTEIN BIOENG
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