Formation of ChromoTek

ChromoTek starts as a spin-off from LMU Munich

Although researches soon after discovery realized the opportunities of alpaca single domain antibodies, the actual development of research reagents proved to be tricky. As probably every scientist has experienced, the devil is in the details. After many experiments, researchers at the LMU eventually succeeded in generating extremely efficient tools for (co-) immunoprecipitation, which were called Nano-Traps. The first Nano-Trap that was developed was the GFP-Trap®. The GFP-Trap proved to be a very useful tool, since many researchers used GFP as tag in their constructs. The outstanding performance of the Nano-Traps and the high demand of researchers led to the foundation of ChromoTek in 2008. In general, ChromoTek’s Nano-Traps are known for exceptionally efficient IPs with single band purification. Numerous protocols are available for a variety of applications, including capture-MS (also known as AP/MS), chromosome IP (ChIP), ribosome IP (RIP). These go along with an impressive number of scientific papers showing data, which were generated using our Nano-Traps.

 

In the following years, ChromoTek has expanded its product portfolio and developed e.g. the innovative and patented Chromobodies® nanoprobes for live cell imaging. Chromobodies are intracellular Nanobodies for real time analysis of endogenous targets in live cells. They are genetic fusions of Nanobodies to fluorescent proteins, which are stable or transiently transduced/transfected to cells. Hence, Chromobodies allow to track endogenous intracellular targets in live cells in real-time. Instead of looking at still (“fixed”) images, cell biologists can now observe the actions of endogenous proteins in real-time. The discovery of Chromobodies was an enormous achievement, because Nanobodies have several cysteine bridges. Even though, the Chromobodies do fold correctly in the reducing environment of the cell’s cytoplasm and do recognize their target. And what made Chromobodies even more attractive as intracellular visualization probes was the fact that they often show very fast on & off kinetics, thus reducing a potential (negative) impact on the cellular function of the target.

ChromoTek has also successfully developed VHHs against linear peptides. These can be used for IP and affinity purification of peptide-tagged fusion proteins. In contrast to the Nanobodies’ preference for 3-dimensional epitopes, which led to the expectation that only properly folded proteins are bound by VHHs, the ChromoTek team developed in 2016 an anti-Myc-tag Nanobody. When coupled to beads, this Myc-Trap can be used for immunoprecipitation and affinity purification of Myc-tagged proteins. In 2018, the novel Spot-Nanobody and short linear Spot-peptide-tag for multiple capture and detection applications was eventually launched. The ChromoTek Spot-system is the first peptide-tag specific Nanobody for universal capture & detection applications. It comprises Spot-Tag®, an inert 12 amino acid peptide-tag, and the universal, rugged Spot-Nanobody that specifically binds to Spot-tagged proteins with high affinity. Spot’s broad applicability ranges from immunofluorescence, super resolution microscopy and Western blot, when Spot-Label, the Spot Nanobody conjugated to fluorophores VHHs, is used to detect Spot-tagged proteins, to immunoprecipitation, affinity purification and additional capture applications, when Spot-Trap, the Spot-VHH coupled to (magnetic) agarose beads, is applied to capture Spot-fusion proteins.

Furthermore, ChromoTek has developed alpaca single domain antibodies for the detection of immunoglobulins in a subtype specific manner. Here, ChromoTek has applied the advantages of Nanobodies to create the product group Nano-Secondaries. These are characterized and validated according to suggestions of the current discussion about quality, validation and traceability of primary antibodies (Getting to reproducible antibodies: the rationale for sequenced recombinant characterized reagents, Bradbury Plückthun (2015), Reproducibility crisis: Blame it on the antibodies, Baker (2015), A proposal for validation of antibodies, Uhlen et al (2016) to name a few).

For more details about ChromoTek’s history, click here.