Endogenous auto-fluorescent biological markers for assessing a biological parameter of a cell

Abstract

The present application relates to the use of endogenous fluorescent biological markers to determine a biological parameter of a cell in a liquid. The methods provided herein provide accurate results in a relatively short amount of time and can be used to monitor and optimize cell culture online as well as determining the presence of a cellular contamination in a cell suspension.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority from U.S. provisional patent applications 61 / 106,176 and 61 / 106,181 both filed on Oct. 17, 2009 and both herewith incorporated by reference in their entirety.BACKGROUND[0002]In order to control and optimize a cell culture, different parameters (growth curves, consumption / depletion of nutrients, production / accumulation of by products (which are usually toxic), determination of physiological state) must be determined at various stages. However, some parameters of a cell culture are very difficult to quantify because there is only a limited number of methods for measuring them during laboratory experimentation or industrial production. Usually, samples of the cell cultures are analyzed with conventional techniques (filtration, drying, cell counting, HPLC). These methods are generally lengthy and costly processes and cannot be performed in real time. Consequently, in industrial settings, cells are generally cu...

AI Technical Summary

Benefits of technology

"The patent text describes a method for determining biological parameters of a cell using an endogenous auto-fluorescent biological marker. The method involves quantifying the fluorescent signal associated with the marker and estimating the biomass concentration or cellular concentration of the cell based on the fluorescent signal. The endogenous auto-fluorescent biological marker can be associated with tryptamine, riboflavin, FAD, or a combination of riboflavin and FAD. The method can be used in a liquid medium containing the cell, and the fluorescent signal can be filtered or suspended in a neutral liquid prior to quantification. The biological parameters that can be determined include biomass concentration, cellular concentration, and cellular proliferation rate."

Problems solved by technology

However, some parameters of a cell culture are very difficult to quantify because there is only a limited number of methods for measuring them during laboratory experimentation or industrial production.

These methods are generally lengthy and costly processes and cannot be performed in real time.

This strategy does not accommodate real time optimization of cell cultures and results in important economic loss.

However, it is virtually impossible to determine in real time other important parameters of the cell culture such as cellular proliferation, physiological state, consumption of nutrients, production of a by-product, etc

Even though the use of endogenous fluorescence to determine the status of a cell culture has proved to be difficult, some research teams have published their efforts toward the understanding of this subject.

However, Hisiger and Jolicoeur also add that “the presence of some unidentified fluorescence signals that are overlapping the ones of interest are limiting the applicability and the reliability of this type of probe”.

For example, it does not provide information about the metabolic behavior of the cells in culture (ex.

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