New highly sensitive sandwich ELISA system for soluble endoglin quantification in different biological fluids. / Smirnov, Ilya Valerevich; Gryazeva, Irina Vladimirovna; Vasileva, Margarita Yurevna; Krutetskaia, Irina Yurevna; Shashkova, Olga Alexandrovna; Samoylovich, Marina Platonovna; Stolbovaya, Anastasia Yurevna; Solodovnikova, Nellya Grigorevna; Zazerskaya, Irina Evegnevna; Sokolov, Dmitriy Igorevich; Selkov, Sergey Alexeevich; Klimovich, Vladimir Borisovich.
In: Scandinavian Journal of Clinical and Laboratory Investigation, Vol. 78, No. 6, 18.08.2018, p. 515-523.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - New highly sensitive sandwich ELISA system for soluble endoglin quantification in different biological fluids
AU - Smirnov, Ilya Valerevich
AU - Gryazeva, Irina Vladimirovna
AU - Vasileva, Margarita Yurevna
AU - Krutetskaia, Irina Yurevna
AU - Shashkova, Olga Alexandrovna
AU - Samoylovich, Marina Platonovna
AU - Stolbovaya, Anastasia Yurevna
AU - Solodovnikova, Nellya Grigorevna
AU - Zazerskaya, Irina Evegnevna
AU - Sokolov, Dmitriy Igorevich
AU - Selkov, Sergey Alexeevich
AU - Klimovich, Vladimir Borisovich
N1 - Publisher Copyright: © 2018, © 2018 Medisinsk Fysiologisk Forenings Forlag (MFFF).
PY - 2018/8/18
Y1 - 2018/8/18
N2 - Soluble endoglin (sEng) is a fragment of a membrane-associated receptor (CD105) expressed on endothelial cells, mesenchymal stem cells and trophoblast cells. It is considered as a regulatory factor involved in the development of preeclampsia (PE) and cancer-associated neo-angiogenesis. The purpose of this study was to describe a new sandwich ELISA for sEng quantification. In contrast to three commercial kits, the new ELISA was able to quantify sEng not only in blood plasma and cell culture supernatants, but also in urine and cerebrospinal fluid. The assay detected up to two orders of magnitude higher antigen levels than did the commercial kits. Using Western blot assay followed by SDS-PAGE or Blue Native PAGE, we demonstrated a heterogeneous nature of sEng molecules. Antigen heterogeneity is considered as a factor contributing to the pronounced differences in its content estimations by different ELISAs. We obtained evidence indicating that the assay was capable of detecting heterogenious sEng molecules. The new ELISA was validated as a quick, specific and accurate method for sEng quantification. Despite the differences in antigen content estimates, the assay had similar diagnostic performance as widely used commercial kit for the detection of severe PE in pregnant women based on plasma sEng contents. Moreover, the new assay was able to delineate diseased patients based on antigen levels in urine. Therefore, the new ELISA is a potentially valuable tool for in vitro and clinical studies.
AB - Soluble endoglin (sEng) is a fragment of a membrane-associated receptor (CD105) expressed on endothelial cells, mesenchymal stem cells and trophoblast cells. It is considered as a regulatory factor involved in the development of preeclampsia (PE) and cancer-associated neo-angiogenesis. The purpose of this study was to describe a new sandwich ELISA for sEng quantification. In contrast to three commercial kits, the new ELISA was able to quantify sEng not only in blood plasma and cell culture supernatants, but also in urine and cerebrospinal fluid. The assay detected up to two orders of magnitude higher antigen levels than did the commercial kits. Using Western blot assay followed by SDS-PAGE or Blue Native PAGE, we demonstrated a heterogeneous nature of sEng molecules. Antigen heterogeneity is considered as a factor contributing to the pronounced differences in its content estimations by different ELISAs. We obtained evidence indicating that the assay was capable of detecting heterogenious sEng molecules. The new ELISA was validated as a quick, specific and accurate method for sEng quantification. Despite the differences in antigen content estimates, the assay had similar diagnostic performance as widely used commercial kit for the detection of severe PE in pregnant women based on plasma sEng contents. Moreover, the new assay was able to delineate diseased patients based on antigen levels in urine. Therefore, the new ELISA is a potentially valuable tool for in vitro and clinical studies.
KW - ELISA
KW - monoclonal antibody
KW - preeclampsia
KW - protein heterogeneity
KW - Soluble endoglin
UR - http://www.scopus.com/inward/record.url?scp=85054291486&partnerID=8YFLogxK
U2 - 10.1080/00365513.2018.1516892
DO - 10.1080/00365513.2018.1516892
M3 - Article
C2 - 30270756
AN - SCOPUS:85054291486
VL - 78
SP - 515
EP - 523
JO - Scandinavian Journal of Clinical and Laboratory Investigation
JF - Scandinavian Journal of Clinical and Laboratory Investigation
SN - 0036-5513
IS - 6
ER -
ID: 89781430