Second, it is very difficult to favor target conformations with non-covalent ligands because they dissociate from the protein shortly after injection, unless their affinities are extremely high, as was the case for the β2 adrenergic receptor agonist BI-167107 having a dissociation constant of as low as 84 pM ( Rasmussen et al., 2011a). many human membrane transporters) readily unfold upon injection due to the applied adjuvants and the camelid’s high body temperature. First, the target space is limited to comparatively stable proteins, because delicate targets (e.g. CDR3 loops of variable length and orientation create diverse binder shapes, thus permitting an optimal surface-complementarity to antigens.ĭespite of the outstanding track record of camelid nanobodies, there are three major restrictions linked to immunizations. The success of nanobodies is rooted in the simplicity and robustness of the VHH scaffold and in its characteristic variability at the complementarity determining region 3 (CDR3), which is frequently found to penetrate deeply into cavities of membrane protein targets ( Kruse et al., 2013 Rasmussen et al., 2011a). A particularly successful method to generate binders against membrane proteins relies on the immunization of camelids for the pre-enrichment of B-cells that encode target-specific heavy-chain-only antibodies, whose variable domains are called VHH or nanobodies ( Pardon et al., 2014). However, binder selection against this difficult class of target proteins is very challenging ( Dominik et al., 2016 Doshi et al., 2014 Pardon et al., 2014), because membrane proteins tend to be flexible, exhibit only small hydrophilic surfaces, require detergents or lipids to remain folded and are typically obtained only in small amounts. The platform does not require access to animal facilities and builds exclusively on commercially available reagents, thus enabling every lab to rapidly generate binders against challenging membrane proteins.Ĭonformation-specific binders raised against membrane proteins have the ability to manipulate cells directly at the cell surface and are exquisite tools for basic science and drug discovery ( Ahuja et al., 2015 Blanpain et al., 2002 Lee et al., 2013). A robust binder selection cascade combining ribosome and phage display enabled the generation of conformation-selective, high affinity sybodies against an ABC transporter and two previously intractable human SLC transporters, GlyT1 and ENT1. To target the limited hydrophilic surfaces of membrane proteins, we designed three sybody libraries that exhibit different shapes and moderate hydrophobicity of the randomized surface.

To overcome this key limitation, we developed an in vitro selection platform based on synthetic single domain antibodies named sybodies. Single domain antibodies are potent reagents for this purpose, but their generation relies on immunizations, which impedes selections in the presence of ligands typically needed to populate defined conformational states. Used in conjunction with a hyperspectral imaging scanning system, hyperspectral imaging has a vast array of lab-based and machine vision applications, including: food grading and inspection, recycling, and cultural heritage analysis.Mechanistic and structural studies of membrane proteins require their stabilization in specific conformations. Headwall's hyperspectral imaging sensors can also be fully integrated for lab-based use, ideal for building classification spectral libraries and validating field-work. land contamination & chemical detection.These images create a map of organic materials within the survey area and can be used in a variety of remote sensing applications including (but not limited to): Hyperspectral cameras optimised for airborne use can be mounted on an unmanned aerial vehicle (UAV), such as a drone in order to capture large area spectral images. With extensive manufacturer technical support, you’ll also have access to our in-house spectral imaging expertise and we are happy to work with you to develop an application tailored to your needs. Headwall Photonics’ spectral instrumentation is proven to deliver excellent results for end users and OEMs across industrial, commercial and government markets. Analytik are exclusive UK and Ireland suppliers of Headwall Photonics', a market leading manufacturer of hyperspectral imaging sensors and spectral instrumentation based in the US.

Single domain protein scaffold