carterra lsa instrument  (Carterra Inc)

Journal: Biomolecules

Article Title: Real-Time SPR Biosensing to Detect and Characterize Fast Dissociation Rate Binding Interactions Missed by Endpoint Detection and Implications for Off-Target Toxicity Screening

doi: 10.3390/biom15060882

Figure Lengend Snippet: Results from fluorescent-based endpoint detection and SPR-based real-time detection of IVTT-expressed HaloTag fusion proteins captured onto glass or SPOC chip capture slide substrates. ( A ) Graphical depiction of the workflow for producing SPOC biosensors via the Protein NanoFactory system. To the right, an image of the SPOC biosensor surface from within the custom Carterra LSA XT SPR instrument flow cell is shown. The array of spots contain various HaloTag fusion proteins covalently captured to the biosensor surface and the hockey stick-shaped fiducial mark enables orientation of the array and identification of HaloTag protein-containing spots. To assess the extent of HaloTag fusion protein capture on glass, an arrayed glass slide was probed with Antibody #1 and detected with Cy3-labeled goat anti-mouse secondary antibody ( B ). The same slide was again probed with Antibody #2, followed by detection using Cy3-labeled goat anti-rabbit secondary antibody ( C ). After each secondary antibody incubation, the slide was scanned on an InnoScan 9100 microarray imager. Presence of a green fluorescent signal at discrete spots indicates detection of HaloTag fusion proteins by the respective primary antibodies. Only two of the four total subarrays present on the glass slide (see for the full glass slide scans) are shown. Sensorgrams of the binding signal obtained from 355 spots across the biosensor surface during ( D ) Antibody #1 (blue) and ( E ) Antibody #2 (green) injections. Both antibodies were injected at 133.3 nM concentration over the course of a 15 min association phase (t = 0.0–900 s), followed by a 30 min dissociation phase (t = 901–2700 s). Each kinetic trace represents the binding signal observed from a single region of interest (ROI) or ligand selected for analysis on the SPOC array. Individual sensorgrams from eachROI reported here are displayed without overlay in .

Article Snippet: The custom LSA XT instrument (Carterra, Salt Lake City, UT, USA) was primed in this running buffer prior to biosensor docking.

Techniques: Labeling, Incubation, Microarray, Binding Assay, Injection, Concentration Assay

Journal: Biomolecules

Article Title: Real-Time SPR Biosensing to Detect and Characterize Fast Dissociation Rate Binding Interactions Missed by Endpoint Detection and Implications for Off-Target Toxicity Screening

doi: 10.3390/biom15060882

Figure Lengend Snippet: Results from fluorescent-based endpoint detection and SPR-based real-time detection of IVTT-expressed HaloTag fusion proteins captured onto glass or SPOC chip capture slide substrates. ( A ) Graphical depiction of the workflow for producing SPOC biosensors via the Protein NanoFactory system. To the right, an image of the SPOC biosensor surface from within the custom Carterra LSA XT SPR instrument flow cell is shown. The array of spots contain various HaloTag fusion proteins covalently captured to the biosensor surface and the hockey stick-shaped fiducial mark enables orientation of the array and identification of HaloTag protein-containing spots. To assess the extent of HaloTag fusion protein capture on glass, an arrayed glass slide was probed with Antibody #1 and detected with Cy3-labeled goat anti-mouse secondary antibody ( B ). The same slide was again probed with Antibody #2, followed by detection using Cy3-labeled goat anti-rabbit secondary antibody ( C ). After each secondary antibody incubation, the slide was scanned on an InnoScan 9100 microarray imager. Presence of a green fluorescent signal at discrete spots indicates detection of HaloTag fusion proteins by the respective primary antibodies. Only two of the four total subarrays present on the glass slide (see for the full glass slide scans) are shown. Sensorgrams of the binding signal obtained from 355 spots across the biosensor surface during ( D ) Antibody #1 (blue) and ( E ) Antibody #2 (green) injections. Both antibodies were injected at 133.3 nM concentration over the course of a 15 min association phase (t = 0.0–900 s), followed by a 30 min dissociation phase (t = 901–2700 s). Each kinetic trace represents the binding signal observed from a single region of interest (ROI) or ligand selected for analysis on the SPOC array. Individual sensorgrams from eachROI reported here are displayed without overlay in .

Article Snippet: Furthermore, we highlight how SPOC technology enhances the multiplex capacity of SPR screening, yielding up to ~864 protein ligand spots in our custom LSA XT Carterra instrument (a ~2.2-fold increase from the standard 384 commercial instrument capacity).

Techniques: Labeling, Incubation, Microarray, Binding Assay, Injection, Concentration Assay