A radioligand binding assay for the insulin-like growth factor 2 receptor

0531943 - ÚOCHB 2021 RIV US eng J - Journal Article
Potalitsyn, Pavlo - Selicharová, Irena - Sršeň, Kryštof - Radosavljević, Jelena - Marek, Aleš - Nováková, Kateřina - Jiráček, Jiří - Žáková, Lenka
A radioligand binding assay for the insulin-like growth factor 2 receptor.
PLoS ONE. Roč. 15, č. 9 (2020), č. článku e0238393. ISSN 1932-6203. E-ISSN 1932-6203
R&D Projects: GA ČR(CZ) GA19-14069S; GA MŠMT(CZ) EF16_019/0000729
Institutional support: RVO:61388963
Keywords : IGF2 * radiologand * binding assay * insulin * iBodies * his-tag
OECD category: Biochemistry and molecular biology
Impact factor: 3.240, year: 2020
Method of publishing: Open access
https://doi.org/10.1371/journal.pone.0238393

Here we produced His-tagged domain 11 (D11), an IGF2-binding element of IGF2R, we immobilized it on the solid support through a well-defined sandwich, consisting of neutravidin, biotin and synthetic anti-His-tag antibodies. Next, we prepared specifically radiolabeled [125I]-monoiodotyrosyl-Tyr2-IGF2 and optimized a sensitive and robust competitive radioligand binding assay for determination of the nanomolar binding affinities of hormones for D11 of IGF2. The assay will be helpful for the characterization of new IGF2 mutants to study the functions of IGF2R and the development of new compounds for the treatment of neurological disorders.
Permanent Link: http://hdl.handle.net/11104/0310572