Recombinant Protein Production Service
01.
Recombinant protein production steps
The production of recombinant proteins involves different steps before obtaining the desired product. See all the summarized steps below to have a better understanding of the process.
Step 01.
Genetic design and strain engineering
Design of high-performance strains to overexpress proteins has a very important role in biochemical production. Genetic design and strain engineering allows to optimize recombinant protein expression in Pichia pastoris/Komagataella phaffii or in Kluyveromyces lactis to obtain higher expression yields. This enhancement can be carried out either in chromosomic DNA or in plasmidic DNA. We are experts in optimizing expression yeast strains with selected characteristics.
Step 02.
Lab-scale culture
Protein expression can be carried out at different scales, according to its expression and the amount of protein required. Lab-scale culture implies the expression in flasks. This method is used to optimize recombinant protein expression since it allows to evaluate the environmental effects of protein expression in a homogeneous media. Our expertise includes the expression optimization at lab-scale.
Step 03.
Bioprocess development & scale-up
Large-scale protein expression becomes one of the most important parts at industrial level. Before expressing large volumes, a scale-up expression from flasks to bioreactor format is necessary to maintain the quality of the results. Additionally, an optimization of the bioprocess is also necessary to further increase the protein expression. Our service involves protein expression and/or its optimization in a bioreactor.
Step 04.
Purification and characterisation
Proteins are expressed in yeast and therefore, other proteins are also present in the mixture. Purification is the removal of protein contaminants through chromatographic methods to obtain the protein of interest isolated. Once the protein is pure, characterization consists in measure its concentration, purity and enzymatic activity. Our technical know-how includes either the purification by different techniques according to the protein properties or its characterization.
Step 05.
Freeze-drying and preservation
The ideal media for proteins is an aqueous media. However, when they are purified, they may need other substances besides water. Additionally, for long-term storage a dehydrated form of the protein is more convenient since it increases the half-life and reduces weight and volume. Lyophilization, also known as freeze-drying, is a process in which water is removed from the sample without using heat. At Levprot, we have a SP VirTis Genesis Pilot Freeze Dryer, which allow us to offer a lyophilization service and its optimization.
