Production of antibodies for therapeutic use on a large scale for pharma industry

Chromatography is a technique that allows the separation, identification, and in some cases the analysis of the different chemical compounds that make up a complex mixture. 

Briefly, the basis of this technique consists in making the mixture, contained in a substance (mobile phase), pass through a solid matrix that is coupled to the system (stationary phase). Thus, depending on the chemical composition of the phases, there will be certain molecules that have a greater affinity for the mobile phase and others that have a greater affinity for the stationary phase. The former, therefore, will move through the system at a higher speed than the latter, thus allowing their separation and detection.

Affinity chromatography, what does it involve?

There are different types of chromatography, with different applications in different fields of study. One of the most widely used types is affinity chromatography, which is generally used for the purification of enzymes, hormones, antibodies, nucleic acids, and specific proteins. 

Briefly, for this type of chromatography, a solution containing the molecule of interest (mobile phase) is passed through a column made up of a solid matrix coupled to a specific ligand (stationary phase). The molecule of interest will recognize the ligand and bind to it by affinity, being retained on the column to be eluted later.

One of the applications of affinity chromatography is the large-scale production of antibodies for therapeutic use, a process that is increasingly widespread in the pharmaceutical industry due to the good results obtained with this type of therapy in various diseases, such as autoimmune diseases or cancer.

This process consists of 4 steps

1. Design and amplification of the DNA vector, which contains the information that codes for the antibody of interest to produce.
2. Transfection of cell lines, so that the DNA is expressed.
3. Large-scale production of the antibody in cell cultures.
4. Purification of the antibody by affinity chromatography.

In greater detail, for step 3, the transfected cells are cultivated in 100mL of culture solution and in optimal conditions for their growth. When cell viability is less than 80%, the cultures will be centrifuged at 3,000 G for 15 minutes, after which the culture supernatant, which must contain the antibodies produced by the cells, as well as other molecules that excreted by the cells during their growth, is collected and filtered. 

This supernatant is stored cold until the purification of the antibodies is carried out by affinity chromatography, which would be step 4.

Affinity chromatography is a particularly important technique in the pharmaceutical industry, as it allows the production of antibodies for therapeutic use on a large scale. 

For this technique to be successful, an intermediate centrifugation step must be performed with an equipment like the universal centrifuge Digicen 21 and its rotor for work with 100 ml samples that can be programmed from 100 to more than 4,000 g.