Multiplexed assays to determine the selectivity of anti-GPCR antibodies
type: Dataset
description: Description of study and method This dataset contains measurement values from a multiplexed screen of antibodies (Abs) to determine their selective recognition of G protein-coupled receptors (GPCRs). Therapeutic antibodies are being developed to modulate GPCR function. However, validating the selectivity of anti-GPCR Abs is challenging due to sequence similarities of individual receptors within GPCR subfamilies. Here we present data from multiplexed immunoassay to test >400 anti-GPCR Abs from the Human Protein Atlas targeting a customized library of 215 expressed and solubilized GPCRs representing all GPCR subfamilies. The GPCRs were conjugated with 1D4 and FLAG epitope tags (common protein tags for Ab binding), were overexpressed in Expi293F cells and solubilized for the assay. The Abs were tested using the suspension bead array (SBA) technology from Luminex. The technology employs color-coded beads with Abs that bind the target (for capture) and fluorescently labelled Abs that bind the 1D4 epitope tag (for detection). The resulting output data are the median fluorescence intensity (MFI) values for each sample and Ab. MFI represents a relative measurement that allows for comparison within Abs but not between. The dataset also contains Z-scores and robust Z-scores for visualization and to provide a similar scale for all Abs. For Ab validation, Abs targeting GPCRs were used for capture. To verify that GPCRs were expressed at all, Abs targeting the FLAG tag were used for capture. Each anti-GPCR Ab was evaluated against its target GPCR and phenotypically closely related GPCRs by using a population density-based threshold to map the on-target and off-target binding of the antibody. The files contain the following data and information - abval_dat.csv: Sample information and Luminex measurements (MFI, Z-scores and robust Z-scores) for samples used in the antibody validation screen (GPCR capture, 1D4 detection). - expr_dat.csv: Sample information and Luminex measurements f…
creative work status: completed
  type: DefinedTerm
  in defined term set: http://publications.europa.eu/resource/authority/dataset-status
  term code: COMPLETED
creator: Leo Dahl
  type: Person
  family name: Dahl
  given name: Leo
creator: Ilana B. Kotliar
  type: Person
  family name: Kotliar
  given name: Ilana B.
creator: Annika Bendes
  type: Person
  family name: Bendes
  given name: Annika
creator: Tea Dodig-Crnkovic
  type: Person
  affiliation: Science for Life Laboratory
  family name: Dodig-Crnkovic
  given name: Tea
creator: Samuel Fromm
  type: Person
  family name: Fromm
  given name: Samuel
creator: Arne Elofsson
creator: Mathias Uhlén
creator: Thomas P. Sakmar
  type: Person
  family name: Sakmar
  given name: Thomas P.
creator: Jochen Schwenk
date modified: 2023-06-02
date published: 2023-02-15
keywords: Medical and Health Sciences
  type: DefinedTerm
  term code: 3
keywords: Biochemistry
  type: DefinedTerm
  term code: 10408
keywords: Biological Sciences
  type: DefinedTerm
  term code: 106
keywords: Receptors and membrane biology
keywords: Analytical biochemistry
keywords: Proteomics and intermolecular interactions (excl. medical proteomics)
license: https://creativecommons.org/licenses/by/4.0/
publisher: Royal Institute of Technology
version: 0

PropertyValue
property ID: DOI
value: 10.17044/SCILIFELAB.21907995

Science for Life Laboratory
type: Organization

Arne Elofsson
type: Person
affiliation: Science for Life Laboratory
family name: Elofsson
given name: Arne

Mathias Uhlén
type: Person
family name: Uhlén
given name: Mathias

Jochen Schwenk
type: Person
affiliation: Science for Life Laboratory
family name: Schwenk
given name: Jochen

Royal Institute of Technology
type: Organization