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A method of determining or predicting a characteristic of a cell

A cell and reference cell technology, which can be applied in the methods of supporting/immobilizing microorganisms, methods of stress-stimulating the growth of microorganisms, biochemical equipment and methods, etc., and can solve problems such as time-consuming and labor-intensive

Active Publication Date: 2016-04-20
VALITACELL
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  • A method of determining or predicting a characteristic of a cell
  • A method of determining or predicting a characteristic of a cell
  • A method of determining or predicting a characteristic of a cell

Examples

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example 1

[0155] Example 1: Identifying Cells Using Stress Response Fingerprints

[0156] Each cell line (CHO-A to CHO-J) was revived from frozen dormancy in two vials. These cell lines were cultured in their recommended growth medium until the growth of each vial stabilized (approximately 4 consecutive subcultures after thawing). The two vials were cultured separately throughout. Using two vials, "bottle-to-vial" variation can be estimated. On day 3 after subculture, two vials of each cell line were inoculated on chemical plates ( figure 1 ). Essentially, each vial of cell lines was grown for 3 days on plates containing the above chemicals. Cells from each vial were seeded on 2 plates, ie 4 plates per cell line.

[0157] After this, the growth level of the cells in the plates was determined (ie in the presence and absence of the aforementioned chemicals) using the "PrestoBlue" assay described above. In this way, the cell-specific growth fingerprint of each CHO cell line can be...

example 2

[0179] Example 2 – Predicting fed-batch performance (IVCD50) of a panel of clonal cells

[0180] A set of 20 clonal cell lines with known fed-batch performance indicators (ie, IVCD50) were self-prepared. Each clonal cell line was grown for 3 days in multi-well plates containing the above chemicals. After this, the level of growth of the cells in the multi-well plate (ie in the presence and absence of the above chemical substances) was determined using the "" assay described above. This allows the identification of the cell-specific growth fingerprint of each clonal cell line. Here, the growth fingerprint was defined as the level of growth of cells in each microenvironment relative to the growth of cells in control wells (ie wells containing medium only).

[0181] An example of a typical growth fingerprint of a clone is figure 2 shown.

[0182] Using the information of these chemical fingerprints, a multivariate linear model was established with the response to each sing...

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Abstract

A method of determining or predicting a characteristic of a query cell, for example the identity of the query cell, is described. The method comprises the steps of incubating the query cell with a plurality of chemical cell stressor molecules, determining the growth response of the query cell in the presence of each of the at least three chemical cell stressor molecules to generate a query cell-specific growth response fingerprint, and comparing the query cell- specific growth response fingerprint with one or more reference cell-specific growth response fingerprints corresponding to one or more cells having known characteristics. The characteristic of the cell (for example the identity of the cell) may be determined based on the level of correlation between the query cell-specific growth response fingerprint and the one or more reference cell-specific growth response fingerprints.

Description

technical field [0001] The present invention relates to methods of determining or predicting characteristics of cells. In particular, the invention relates to a method of determining cell identity or predicting cell fed-batch performance. Background technique [0002] Many biopharmaceuticals are produced using genetically modified single-celled organisms, called producer cells. A well-established system for producing biopharmaceuticals using mammalian producer cells has been established. The first step involves obtaining highly productive cell lines suitable for large-scale production, a process that begins with transfection, cloning, and then characterizing a large number of cell lines. The process is costly, labor-intensive and time-consuming, lasting up to 12 months. Once transfected with the gene of interest and applying selective pressure using a selective expression system, the next steps involve the selection and isolation of cells with acceptable growth and recomb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G05B17/02G05B13/04C12M1/36C12M1/34
CPCG01N33/5014G01N33/5005C12M23/12C12M25/06C12M35/08C12M41/46
Inventor B·汤普森D·詹姆斯S·L·戴维斯
Owner VALITACELL
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