Journal: mAbs

Article Title: Generation of robust bispecific antibodies through fusion of single-domain antibodies on IgG scaffolds: a comprehensive comparison of formats

doi: 10.1080/19420862.2023.2189432

Figure Lengend Snippet: Antibody binding to PD-L1 and HER2 using FIDA. (a) Assessment of bispecific binding functionality through changes in apparent Rh in response to addition of fluorescent PD-L1-DY490 ± unlabeled HER2 antigens in solution (b) Assessment of bispecific binding functionality through changes in apparent Rh in response to addition of fluorescent HER2-DY490 ± unlabeled PD-L1 antigens in solution. (c) Antibody titration curves investigating binding in mono- and bispecific binding environments. The binding curves were generated by titrating antibody against the fluorescently labeled primary antigen ± an unlabeled secondary antigen. Since the secondary antigen does not carry any fluorescence, it can only affect the signal indirectly through complexation with the bsAb, which is in itself binding the primary antigen. This is to test if the titration curves change in response to addition of the unlabeled secondary antigen. The coloring scheme is the same as in (a) and (b).

Article Snippet: Rh values were obtained by Taylorgrams to FIDA Software v2.3 (Fida Biosystems) with a Taylorgram fraction of 75%.

Techniques: Binding Assay, Titration, Generated, Labeling, Fluorescence

Journal: mAbs

Article Title: Generation of robust bispecific antibodies through fusion of single-domain antibodies on IgG scaffolds: a comprehensive comparison of formats

doi: 10.1080/19420862.2023.2189432

Figure Lengend Snippet: Affinity binding constants and the related goodness-of-fit from antibody titration curves using FIDA. The titration curves were generated by titrating antibodies against a fluorescently labeled antigen ± the unlabeled antigen. The goodness-of-fit is evaluated using R 2 and root mean squared error (RMSE). Affinity values in brackets marked with an asterisk indicate affinity values previously reported elsewhere. 20 , 21 , 33 , 34 .

Article Snippet: Rh values were obtained by Taylorgrams to FIDA Software v2.3 (Fida Biosystems) with a Taylorgram fraction of 75%.

Techniques: Binding Assay, Titration, Generated, Labeling

Journal: The Journal of Physical Chemistry. B

Article Title: Two Receptor Binding Strategy of SARS-CoV-2 Is Mediated by Both the N-Terminal and Receptor-Binding Spike Domain

doi: 10.1021/acs.jpcb.3c06258

Figure Lengend Snippet: FIDA of SARS-CoV-2 spike S1 segment binding to SA containing glycolipids. (a) Schematic of how complex formation by fluorescently labeled S1 will affect the Taylorgrams of FIDA measurements. A single species that includes a fluorescent label will give rise to a single Gaussian line shape, with a Gaussian width σ that increases with an increasing hydrodynamic radius. (b) Taylorgrams recorded for pure 100 nM spike S1 protein, with 200 μM DPPC vesicles, and with 1:9 GM1:DPPC vesicles, partly composed of the SA containing GM1 glycolipid. In the case of the glycolipid-containing vesicles, a double Gaussian line shape is observed, indicating the presence of both free S1 species and S1-vesicle complexes, as opposed to the single Gaussian lineshapes observed for the pure S1 for in the presence of the DPPC vesicles that do not contain glycolipids, indicating the absence of S1-binding in that case. (c) The relative number of bound species for the S1 segments of SARS-CoV, SARS-CoV-2, and MERS-CoV bound to (1:9)-GM1:DPPC and -GM3:DPPC lipid vesicles (lipid concentration in monomer units). Lines show best fit to eq . (d) Estimated dissociation constants (K D s).

Article Snippet: Finally, the injected indicator sample is then flowed toward the detection point with the vesicle sample at 50 mbar for 20 min. All samples were performed in duplicate, and the Taylorgrams were processed using the FIDA data analysis software (Fida Biosystems ApS, Copenhagen, Denmark).

Techniques: Binding Assay, Labeling, Concentration Assay