Dataset DOI: 10.5061/dryad.f4qrfj77j

Description of the data and file structure

Dataset DOI: 10.5061/dryad.f4qrfj77j

In this study, we present a phage-displayed synthetic library for nanobody discovery. To validate this approach, we screened nanobodies targeting eight Drosophila secreted proteins. The nanobodies identified were validated by ELISA, immunostaining, and immunoblotting. This dataset contains all the source data in the manuscript, including polyclonal ELISA data, monoclonal ELISA data, immunostaining data, and Western Blotting data.

Files and variables

File: Source_data.zip

Description: This dataset contains 7 file folders. The file folders are named after their corresponding figures in the manuscript. In each file folder, the files are named after their corresponding panels in the figures.

*Figure 2-source data: Nanobody screen for CARPB. Figure 2B polyclonal ELISA source data and Figure 2C monoclonal ELISA source data for CARPB nanobodies. In Figure 2B polyclonal ELISA source data, the columns represent the polyclonal phages from the first round, second round, and third round screen. The rows represent the decreasing amount of mCherry-hIgG and CARPB-hIgG coated across different rows. The values are obsorbence at 450 nm (OD450). The intensity of green color represents the ELISA signal strength. In Figure 2C monoclonal ELISA source data, the rows represent different wells and the columns represent the monoclonal phages reacted with mCherry-hIgG and CARPB-hIgG. 

*Figure 3-source data: Immunostaining of membrane-tethered CARPB with nanobodies. Figure 3A immunostaining source data and Figure 3B immunostaining source data for immunostaining with CARPB nanobodies. The .jpg file names represent the antigen (CARPB-GPI), the monoclonal phages and channels (bf, bright field; GFP, transfection marker for antigen CARPB-GPI expressing cells; M13/VHH, phage/nanobody immunostaining signal)

*Figure 4-source data: Immunostaining and immunoblotting with NbMip-4G nanobody. Figure 4A immunostaining source data and Figure 4B Western Bloting source data for NbMip-4G nanobody. In Figure 4A immunostaining source data, the .jpg file names represent the fly genotype (Mex-Gal4w1118 or Mip RNAiv106076) and channels (DAPI, NbMip-4G immunostaining signal and merged)

*Figure 2-figure supplement 1-source data: Nanobody screen for Nesfatin-1. Figure 2-figure supplement 1A polyclonal ELISA source data, Figure 2-figure supplement 1B monoclonal ELISA source data, Figure 2-figure supplement 1C immunostaining source data, Figure 2-figure supplement 1D immunostaining source data for Nesfatin-1 nanobody screen.

*Figure 2-figure supplement 2-source data: Nanobody screen for Mip. Figure 2-figure supplement 2A polyclonal ELISA source data, Figure 2-figure supplement 2B monoclonal ELISA source data, Figure 2-figure supplement 2C immunostaining source data, Figure 2-figure supplement 2D immunostaining source data for Mip nanobody screen.

*Figure 2-figure supplement 3-source data: Nanobody screen for CG9849. Figure 2-figure supplement 3A polyclonal ELISA source data, Figure 2-figure supplement 3B monoclonal ELISA source data, Figure 2-figure supplement 3C immunostaining source data, Figure 2-figure supplement 3D immunostaining source data for CG9849 nanobody screen.

*Figure 2-figure supplement 4-source data: Nanobody screen for Amn. Figure 2-figure supplement 4A polyclonal ELISA source data, Figure 2-figure supplement 4B monoclonal ELISA source data for Amn nanobody screen.

*Figure 2-figure supplement 5-source data: Failed nanobody screens for Sgsf, Miple1, and CG13965. Figure 2-figure supplement 5A monoclonal ELISA source data for Sgsf nanobody screen, Figure 2-figure supplement 5B monoclonal ELISA source data for Miple1 nanobody screen, Figure 2-figure supplement 5C monoclonal ELISA source data for CG13965 nanobody screen.

Code/software

None

Access information

Other publicly accessible locations of the data:

• None

Data was derived from the following sources:

• None