Affinity maturation of antibody against membrane-bound GPCR molecules
Ontology highlight
ABSTRACT:
PROVIDER: PRJEB32492 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:G protein-coupled receptors (GPCRs), also known as seven-transmembrane domain receptors, are among the most important targets against which many small molecule drugs have been developed. However, only two antibody drugs targeting GPCRs have been approved for clinical use although many antibody drugs against non-GPCR protein targets have been successfully developed for various disease indications. One of the challenges for developing anti-GPCR drugs is the high difficulty to perform affinity maturation due to their insolubility in aqueous solutions. To address this issue, CHO cell display libraries of single-chain variable fragments (scFvs) and full-length antibodies were maturated directly against vesicle probes prepared from CHO cells displaying the endothelin A receptor (ETaR) GPCR. The probe in the vesicle form ensures the physiological conformation and functional activity of the protein and avoids issues with membrane protein insolubility. The size of the vesicle had a clear effect on protein-ligand interaction; we used small-sized vesicles with low expression levels of GPCRs for the affinity maturation. Four rounds of affinity maturation combining vesicles as probes with the CHO cell display platform improved affinity by 13.58-fold for scFvs and 5.05-fold for full-length antibodies. We expect that this method will not only be used for the affinity maturation of antibodies against GPCRs but will also be used to mature antibodies for other types of proteins where the conformation/activity of which depends on the proper membrane environment.
| S-EPMC6719327 | biostudies-literature
Project description:Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. mRNA species bound to membrane-associated polysomes were separated from other mRNAs by sedimentation equilibrium or sedimentation velocity. The distribution of individual transcripts in the 'membrane-bound' and 'cytosolic' fractions was quantitated for thousands of genes by hybridization to DNA microarrays. Transcripts known to encode secreted or membrane proteins were enriched in the membrane-bound fractions, whereas those known to encode cytoplasmic proteins were enriched in the fractions containing mRNAs associated with free and cytoplasmic ribosomes. On this basis, we identified over 275 human genes and 285 yeast genes that are likely to encode previously unrecognized secreted or membrane proteins. Computed
2005-08-24 | E-GEOD-3180 | biostudies-arrayexpress
Project description:Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. mRNA species bound to membrane-associated polysomes were separated from other mRNAs by sedimentation equilibrium or sedimentation velocity. The distribution of individual transcripts in the 'membrane-bound' and 'cytosolic' fractions was quantitated for thousands of genes by hybridization to DNA microarrays. Transcripts known to encode secreted or membrane proteins were enriched in the membrane-bound fractions, whereas those known to encode cytoplasmic proteins were enriched in the fractions containing mRNAs associated with free and cytoplasmic ribosomes. On this basis, we identified over 275 human genes and 285 yeast genes that are likely to encode previously unrecognized secreted or membrane proteins. Keywords: Logical Set
2005-08-24 | GSE3180 | GEO
Project description:RNA of dorsal root ganglia from untreated adult mice (6-8 weeks) was tested for GPCR expression
2015-12-01 | GSE75476 | GEO
Project description:Subcellular location defines GPCR signal transduction
| PRJNA915973 | ENA
Project description:Identification of human tRNA molecules bound to human cytomegalovirus capsid using cryoEM and Next-generation sequencing
| PRJNA702772 | ENA
Project description:Analysis of the effect of G-alpha or GPCR mutation on the Arabidopsis Transcriptome Keywords: Comparitive Genome Hybridization The transcriptomes of wild type and mutants (G-alpha and GPCR) were compared in the current study. The study was carried out on four whole genome oligo arrays. The G-alpha mutant and its corresponding wild type variety were compared on a single array by means of a dual channel experiment (Cy3 and Cy5 labeling). The experiment was repeated on a different array as a flip-dye replicate and the data from both the sets analyzed further.
2010-09-30 | E-GEOD-11435 | biostudies-arrayexpress
Project description:This study shows that chemically and pharmacodynamically distinct agonists acting on the same GPCR can produce indistinguishable cellular responses and that this uniformity is conferred by endosomal signaling The ability of chemically distinct ligands to produce different effects on the same G protein-coupled receptor (GPCR) has interesting therapeutic implications but, if excessively propagated downstream, would introduce biological 'noise' compromising cognate ligand detection We asked if cells have the ability to limit the degree to which chemical diversity imposed at the ligand-GPCR interface is propagated to the downstream signal We carried out an unbiased analysis of the integrated cellular response elicited by two chemically and pharmacodynamically diverse β-adrenoceptor agonists, isoproterenol and salmeterol We show that both ligands generate an identical integrated response, and that this stereotyped output requires endocytosis We further demonstrate that the endosomal β2-AR signal confers uniformity on the downstream response because it is highly sensitive and saturable Based on these findings, we propose that GPCR signaling from endosomes functions as a biological noise filter to enhance reliability of cognate ligand detection
2016-11-30 | GSE87461 | GEO
Project description:In human HCC tumors, matching paracancerous specimens, and normal liver specimens, a targeted gene expression microanalysis was used to screen alterations in GPCR family gene expression.
2017-03-07 | GSE95698 | GEO
Project description:CRISPR/Cas9 genome editing was used to disrupt nearly all the GPCR and neuropeptide genes from C. elegans genome. Multiple genes were disrupted in each strain for the purpose of screening. The genotype is the list of targeted genes
2023-06-02 | E-MTAB-12983 | biostudies-arrayexpress