When considering the costs incurred for a protein structure determination, the first question that arises is whether the protein structure has already been solved with a slight change (inhibitor, mutation) or whether this is the first structure determination of the protein. In that case it is referred to as a de novo structure determination. Here, the effort is usually much higher and thus, the costs are correspondingly higher.
The protein crystallographic determination of a de novo structure starts at around €5,000 and can easily amount to €200,000. This essentially depends on how much effort is required to optimize the crystallization condition and how easily the phases and the cryo condition can be determined.
The subsequent structures are usually in the range of €2,500 to €5,000. This depends on how much experience you have with the protein, how good the model structure is, whether a script is used for the structure solution, what refinement state you need, and the size of the protein (amino acids and molecules in the asymmetric unit).
The de novo structure determination using cryo-EM is much better for predicting the costs. Expect to pay around €100,000 here. The greatest effort lies in determining the right conditions for the production of the grids. For the subsequent structures you should calculate with about 15,000 € to 20,000 €.
Due to the lower costs, it is recommended to start with protein crystallographic structure determination, i.e. commission a crystallization screen and see if the protein crystallizes. Ideally, the crystal can be measured directly at the synchrotron and the structure can be solved. €5,000 is quite realistic here, although it is better to calculate with €10,000 to €15,000, especially if the structure is to be deposited in the protein database.
Cryo-EM is recommended if you want to determine the structure of a membrane protein, if there is only a small amount of protein available, or if no suitable crystallization condition was found in a crystallization screening. However, in addition to the higher costs, cryo-EM has the disadvantage that the resolution is usually much lower, so that no clear statement can be made regarding the orientation of a side chain or the inhibitor. In addition, the structure of proteins with a molecular weight of less than 50 kDa cannot be determined using cryo-EM. Fusing with another protein can possibly lead to success here.
If you do not just want to commission the de novo structure determination of your protein, but want to determine the structure together with a service provider, the following concept can be suitable for you. Through targeted training, we will enable you to understand the service provider's approach and to support him experimentally, so that costs can be reduced, you expand your expertise in the company, and increase your degree of autonomy.
At the beginning, you will complete the theory module (I) of the protein crystallography course to be able to understand the process of de novo structure determination. You order a crystallization screen of your protein and ask the service provider for remote access to the automatically photographed crystallization plates in order to trace the crystal growth. Ideally, a crystal of the screen can be bombarded with X-rays directly on site at the synchrotron and the structure can be solved. A crystallization screen can be carried out quickly and is only associated with low costs.
Usually, the quality of the crystals in a crystallization screen is not suitable for direct data acquisition, so that the crystallization conditions have to be optimized. Module II and module III of the protein crystallography course empowers you to independently reproduce the crystallization condition of the screen in your company and to optimize the crystallization condition. In addition, after completing the course, you will be able to transfer the optimized crystals into liquid nitrogen and send them to the synchrotron so that a dataset can be collected.
After the structure solution, you will receive a coordinate file (pdb file) in addition to the other processing and refinement files. On request, with our special training we will enable you to generate meaningful images from the coordinate file and to analyze the structures yourself.