The ethical practices experimental animals have made the production of antibodies vitro in by the hybridoma culture in the laboratory to gain more ground over traditional production in vivo from ascites fluid.
Despite the more than notable advances in in vitro antibody production techniques, difficulties such as low yield, cell death, contamination of cultures and / or scalability problems, among others, often arise.
In order to solve and optimize these questions, today we bring you this post with some keys for the production of antibodies in vitro .
1.- Optimization Of The Culture Medium
Each hybridoma responds differently to a certain culture medium, which will directly influence cell growth, and by extension, the quality and performance of the antibody produced.
The most important factors to consider when selecting the culture medium for hybridomas are:
- Culture media containing nutrients and additives such as growth factors, growth inhibitors, autocrine factors, cyclic nucleotides, etc. they can increase the production of antibodies in vitro .
- In some cases, one way to stimulate the production of antibodies in vitro is the use of culture media that generate high osmotic pressure .
- The generation of metabolic by-products can be reduced using culture media with low concentrations of glucose and glutamine . In this way cell viability is prolonged, improving performance.
- In vitro antibody production can also be enhanced by amino acid rich culture media .
- To optimize the action of the selected culture medium and thus improve the levels of production of antibodies in vitro , it is necessary to define the appropriate physical-chemical conditions in each case, including pH, temperature, and oxygen and CO2 levels.
2.- Optimization Of The Antibody Production System
Depending on the available resources, the scale of production and the amount of antibody required, different in vitro antibody production systems can be chosen .
Let’s see the characteristics that each of them offers:
- BATCH CULTIVATION
In this case, the nutrients are added only once at the beginning of the cultivation, and each batch is processed independently.
- Features :
- Method of choice for small-scale production of antibodies in vitro .
- The procedure is simple, relatively quick (3 weeks), and inexpensive.
- The yield does not usually exceed 20ug / mL.
- CONTINUOUS CULTIVATION
In this modality, the nutrients are added as they are consumed and it allows working based on continuous cultivation.
- Features :
- By working with higher cell density, this method provides higher performance.
- They use less amount of medium.
- Requires more expensive equipment and more continuous monitoring.
- PERFUSION CULTURE
In this case, the nutrient medium is circulated over the cells through a semipermeable membrane.
- Features :
- Method of choice for large scale in vitro antibody production .
- Requires more investment in equipment.
- The performance obtained is much higher than with any of the other methods, since the cell density achieved is much higher. It can reach up to 1mg / mL.