Extended Data Fig. 5: Oligo-barcoded Anti-VEGF-A binding specificity on nanovials.
From: Associating growth factor secretions and transcriptomes of single cells in nanovials using SEC-seq
a, An anti-VEGF-A antibody (used as detection antibody for VEGF-A secretion in our SEC-seq approach) was conjugated with a 10x Chromium compatible oligo-barcode along with additional sequences necessary for library preparation. The schematic shows the sequence composition of the oligo attached to the VEGF-A detection antibody, along with the 10x Chromium primer sequence which hybridizes to the antibody-derived oligo and adds the unique molecular identifier (UMI) upon reverse transcription. b, (Left) Schematic showing the attachment of recombinant VEGF-A and the detection immunoassay via the oligo-barcoded antibody described in (a) which was quantified with a fluorescently-labelled secondary antibody in nanovials by flow cytometry. (Right) Flow cytometry histograms showing outcome of the experiment on the left with recombinant VEGF-A with and without the oligo-barcoded VEGF-A detection antibody, demonstrating antibody specificity of the detection assay and the validity of the oligo-barcoded VEGF-A detection antibody to the presence of VEGF-A protein. The numbers on the left indicate the mean of both histograms. c, As in (b), except that no biotinylated anti-VEGF-A capture antibody and recombinant VEGF-A was used in the assay. Schematics in created with BioRender.com.
