Derisking your crystallization parameters in 6 weeks Utilizing 1000+ conditions using Broad, Sparse Matrix, Grid and Fine Screenings.
Crystallization of FAB Antigen and Antibody complexes to empower your biologics development.
- FAB (Fragment Antibody Binding) Region Characterization, especially relevant in Immunology and Immuno-Oncology.
- FAB and Antibody complexes to decipher molecular interactions at atomic resolution.
Target Engagement With Small Molecules and Peptides
- Targeted Small Molecule/Peptide- Protein Interactions using Co-crystallization and Soaking Platforms.
- Advance your knowledge with atomic resolution of your ligand in its binding pocket.
Enter your information below for cost-effective de-risking of your targets and the feasibility of crystallization.
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Primary Screening is to establish crystallographic feasibility and conditions to explore and will be followed by optimization screen.
Protein is subjected to 1000+ crystallization conditions. 6 weeks of monitoring is included:
- First week includes daily reporting.
- Following 5 weeks include weekly reporting.
- Final full report at 6 weeks: Lists the hits from the screen that show potential for crystallization and need further refinement.
Frequently Asked Questions
Optimization Screens/Soaking Platforms, Data Collection/Refinement, and Structure Solutions are the additional steps we can take to find the crystal structure.
As with all things in crystallography there are no hard and fast rules, but a good start concentration is 10 mg/mL if it can be achieved while maintaining homogeneity as determined by size exclusion chromatography. We regularly see well-behaving crystals at higher and lower concentrations.
Primary crystallization screening would require about 2-3 mg of purified protein. The protein should be 95-98 % pure as analyzed on Coomassie-blue stained gel. Another 2-3 mg of protein would be required for optimization screens and for growing crystals that will be sent to the synchrotron beamlines for data collection. More is always better!!
The amount of protein that is available directly informs the path of screening. If you have more, you are allowed to go broad and if you have less, you need to screen less. At Ichor, we utilize the FORMULATRIX NT8. This allows the precise setting of nano liter volumes of liquid in a humidified chamber to reduce evaporation. So how much protein do you need? If you assume a concentration of 10 milligrams per milliliter of protein utilized in the crystallization drop and a drop size of 100 nanoliters you will need ~1 microgram per drop. A traditional set up would be about 1,000 conditions which would equal ~1 milligrams. An additional milligram of protein is consumed in characterization and concentration which is why at a minimum, we li recommend 2-3 milligrams
Primary crystallization screening involves incubating the protein under conditions varying different physicochemical parameters such as concentration of precipitants, concentration of salts, pH and temperature. The conditions used are made by identifying conditions that have most frequently produced protein crystals. We generally screen about 800-1000 conditions during our initial primary screen.
Optimization screening involves varying physicochemical parameters around the promising conditions that have been identified from the primary screens. For example, if 20% PEG8000 and 01. M Tris-HCl (pH 7.5) are identified as a missing condition, then optimization screens would be setup varying the PEG 8000 from 10 – 30% and the pH would be varied from 7.0 – 8.0.
No. Our comprehensive capabilities include advanced protein expression and purification in bacteria, yeast, insect, and mammalian cell culture. We specialize in making high quality protein at scale for your biophysical and structural biology needs.
No. We are a flexible crystallography shop where we allow the receipt of client provide samples to be used in structural biology and biophysical work flows. Our team of protein chemists handle your protein and characterize its homogeneity and concentration upon receipt. We typically set up screens within a day receiving client’s protein.
The timeline for a specific screening campaign depends on the amount of protein provided and the handling conditions and crystallographic conditions to make strong crystals. On average a primary screening campaign is 6 weeks. Where the plates are monitored daily. If any hits are found in these conditions, then crystal farms and optimization screens are set up around those conditions to get more crystals.
The short answer is one! On average we like to take batches of approximately 30 crystals per condition. This allows us to optimize the cryoprotectant and have a better chance of finding a clean well diffracting crystal.
A normal 50 kd protein would be about 5 weeks but this can take longer if the protein is larger or if there are a lot of undefined regions in the electron density map.