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EP-4090948-B1 - IR SPECTROSCOPIC CELL CULTURE ANALYSIS

EP4090948B1EP 4090948 B1EP4090948 B1EP 4090948B1EP-4090948-B1

Inventors

  • HEGARTY, MARK
  • BAKER, MATTHEW, J.
  • BUTLER, HOLLY

Dates

Publication Date
20260513
Application Date
20210115

Claims (12)

  1. A method for detecting a status of cells within an in vitro cell culture, the method comprising: providing an ATR-FTIR spectrum from a test sample of a cell medium within an in vitro cell culture; and comparing the ATR-FTIR spectrum with an ATR-FTIR spectrum of a control sample or samples, or a sample or samples obtained at an earlier time point of the cell medium within the in vitro cell culture to the test sample, in order to detect any difference between the test and control or earlier samples, which can be correlated with the status of cells within the test sample obtained from the cell medium within the in vitro cell culture, wherein the status of cells within the cell culture relates to the health of the cells, and wherein the IR spectra are collected at wavenumbers in the region of 400-4000 cm -1 .
  2. An integrated cell culture and ATR-FTIR analysis system comprising: a) in vitro cell culture apparatus for growing cells in culture; b) an ATR-FTIR spectrometer; and c) a sample handling system for obtaining one or more samples of a cell medium within a cell culture within the in vitro cell culture apparatus, the sample handling system comprising a sampler for obtaining a sample of the cell medium within the cell culture and transporting the sample to the ATR-FTIR spectrometer in order that an ATR-FTIR spectrum of the sample or samples may be obtained at wavenumbers in the region of 400-4000 cm -1 ; wherein the system is configured to conduct the method according to claim 1, so as to detect the status of cells within the cell culture, wherein the status of cells within the cell culture relates to the health of the cells.
  3. The method according to claim 1, comprising obtaining the sample or samples from the cell culture manually, in a semi-automated manner, or in a fully automated manner.
  4. The method according to claim 1, the method being implemented and/or controlled by a computer system with integrated software in order to conduct the comparison step and detect any differences between the test sample and control or earlier samples obtained from the cell culture.
  5. The method according to claim 1, wherein the status of the cells is detected as being healthy, or unhealthy.
  6. The method according to claim 5 wherein the unhealthy status of the cells, is due to the cells being infected by a virus or bacteria, or due to the cells having developed an abnormality, optionally wherein the abnormality has occurred due to cell culture conditions, such as a change in pH, oxygen levels, toxic component build up and/or nutrient levels.
  7. The system according to claim 2, wherein the integrated system further comprises the ability to, alert a user, alter and/or halt the cell culturing process in response to the status obtained from the method.
  8. The method according to claim 1, wherein the cell culture comprises cells which produce a product, such as a recombinant protein or the like; or, the cells are the product, optionally wherein the cells are the product and are intended for in vivo and/or in vitro application.
  9. The method according to claim 1, wherein the cells are bacterial or eukaryotic, such as mammalian or other eukaryotic cell types.
  10. The method according to claim 1 wherein the step of comparing the IR spectra is performed using a predictive model, optionally which has been developed by training a database of pre-correlated analyses.
  11. A computer installed with computer software configured to operate the computer to perform the method according to claim 1, optionally wherein the computer incorporates a predictive model derived from one or more pattern recognition algorithms applied to a plurality of pre-correlated ATR-FTIR spectra or processed ATR-FTIR spectra, optionally further wherein the computer is installed with a database for correlating results with known cell status samples and/or to store training set data.
  12. A computer-readable medium containing computer software and optional database, wherein when the software is executed by a computer, it causes the computer to carry out the method of claim 1.

Description

Field of the Invention The present invention relates to methods for performing infrared spectroscopy (IR) analysis on cell cultures, in order to detect any potential abnormalities in cells, which are present in the cell culture. Background of the Invention Fourier Transform Infrared (FTIR) spectroscopy is a technique commonly used in chemical sciences in order to identify discrete vibrations of chemical bonds. This technique uses light generally in the mid-infrared (MIR) region (4000 - 400 cm-1) that is, in the same frequency range as the frequency range of specific chemical bond vibrations. Biological molecules are known to actively vibrate in this range of wavelengths, and thus FTIR spectroscopy lends itself to biological applications. When a biological sample is irradiated with MIR light, some of this energy is absorbed by the sample. The absorption profile of a given sample is representative of the chemical bonds present within a sample, and can be used to characterise complex biological materials. An example of a particular type of analysis using FTIR spectroscopy is in the investigation of proliferative disorders, such as cancer, which are caused by uncontrolled and unregulated cellular proliferation and can, in some cases, lead to the formation of a tumour. Recently a method of diagnosing brain cancer by performing Attenuated Total Reflection - Fourier Transform Infrared (ATR-FTIR) spectroscopic analysis of blood samples has been described in WO 2014/076480). In contrast to conventional ATR-IR (where a sample is placed on a substrate that is then brought into contact with the ATR crystal), the ATR crystal was used as the substrate for the sample. This method provides a point of care and nondestructive diagnostic test. In the field of biological product manufacture using cells, known as bioprocessing, the cells are typically initially grown in small bench-scale batches, before scale-up. Scaling-up of the cell culture and cells within the culture, typically mammalian cells, is both time consuming and costly. Moreover, the biological product which is to be produced is generally of high value and so it is important that the cells which are used to produce the biological product are healthy, in order to produce the highest levels and quality of the biological product. However, the scaling-up process may be carried out over a number of weeks and/or in different increasingly sized bioreactors, before the final full-scale process and bioreactor is achieved. This scaling-up of an upstream process can result in cell abnormalities occurring, such as due to contamination causing bacterial or viral infection, or problems associated with inhibited or inefficient cell growth. The sooner such abnormalities can be detected the better, in terms of process control and cost efficiencies. Moreover, in some instances, the required biological product may only be expressed once the cell culture has been scaled-up to its maximum and in such instances, a user may not know there is any problem with the cells and hence their ability to produce the biological product, until after a great deal of time and expense has been incurred. Relevant prior art is disclosed in US 2011/236922 A1 and US2015/301017 A1. In view of the above, it would be desirable to be able to monitor cells within a cell-culture, typically during any stage of a bioprocess, including upstream and downstream processes, in order to be able to quickly ascertain the health of the cells within the cell culture and whether or not the scaling-up process is proceeding satisfactorily, or not. Summary of the Invention The present teaching is based on studies made by the inventors into IR analysis of cells and/or the cell medium within a cell culture and the inventors' observations that it is possible, from the IR analyses, to detect differences, such as healthy versus unhealthy status, in cells. Thus, in a first aspect, there is provided a method for detecting a status of cells within a cell culture, as claimed in claim 1. Thus, in a further aspect there is provided an integrated cell culture and IR analysis system as claimed in claim 2. In a further aspect there is provided a computer system with integrated software as claimed in claim 11. Such a computer system will typically include a display device in order to provide a user with a result of the comparison. The "status" of cells within the cell culture, relates to the health of the cells. That is, whether or not the cells within the cell culture are considered to be healthy, or unhealthy. Unhealthy cells may be diseased, such as due to infection by a virus or bacteria, for example, or may have developed an abnormality due to the cell culture conditions and/or a mutation occurring in the cells. An abnormality occurring due to cell culture conditions may arise due to, for example, a change in pH, oxygen levels, nutrient levels, and the like. Essentially, any less than optimum conditions may result in changes occurring i