mesЭкстремальная биомедицинаExtreme Medicine3033-89643033-8972Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2023.002mes-84Research ArticleМЕТОДMETHODИдентификация фосфонилированных пептидов с применением МАЛДИ-мишени, функционализированной стеаратом лантанаIdentification of phosphonylated peptides using a MALDI target functionalized with lanthanum stearateБабаковВ. Н.BabakovV. N.

Владимир Николаевич Бабаков 

г.п. Кузьмоловский, ст. Капитолово, корп. 93, Ленинградская обл. 

Vladimir Nikolaevich Babakov 

Kuzmolovsky, st. Kapitolovo, str. 93, Leningradsraja oblast 

babakov@gpech.ruГорбуновА. Ю.GorbunovA. Yu.

Санкт-Петербург

St. Petersburg

ГладчукА. С.GladchukA. S.

Санкт-Петербург

St. Petersburg

КалниняЯ. К.KalninyaYa. K.

Санкт-Петербург

St. Petersburg

ШиловскихВ. В.ShilovskikhV. V.

Санкт-Петербург

St. Petersburg

ТомилинН. В.TomilinN. V.

Санкт-Петербург

St. Petersburg 

СуходоловН. Г.SukhodolovN. G.

Санкт-Петербург

St. Petersburg

РадиловА. С.RadilovA. S.

Санкт-Петербург

St. Petersburg

ПодольскаяЕ. П.PodolskayaE. P.

Санкт-Петербург

St. Petersburg

Научно-исследовательский институт гигиены, профпатологии и экологии человека Федерального медико-биологического агентстваРоссияResearch Institute of Hygiene, Occupational Pathology and Human Ecology of the Federal Medical Biological AgencyRussian FederationНаучно-клинический центр токсикологии имени С. Н. Голикова Федерального медико-биологического агентства ; Санкт-Петербургский государственный университетРоссияGolikov Research Center of Toxicology ; St. Petersburg State UniversityRussian FederationНаучно-клинический центр токсикологии имени С. Н. Голикова Федерального медико-биологического агентстваРоссияGolikov Research Center of ToxicologyRussian FederationСанкт-Петербургский государственный университетРоссияSt. Petersburg State UniversityRussian FederationИнститут аналитического приборостроения Российской академии наук ; Санкт-Петербургский государственный университетРоссияInstitute for Analytical Instrumentation, Russian Academy of Sciences ; St. Petersburg State UniversityRussian FederationНаучно-клинический центр токсикологии имени С. Н. Голикова Федерального медико-биологического агентства ; Институт аналитического приборостроения Российской академии наукРоссияGolikov Research Center of Toxicology ; Institute for Analytical Instrumentation, Russian Academy of SciencesRussian Federation2023241020242512130Copyright © Бабаков В.Н., Горбунов А.Ю., Гладчук А.С., Калниня Я.К., Шиловских В.В., Томилин Н.В., Суходолов Н.Г., Радилов А.С., Подольская Е.П., 20242024Бабаков В.Н., Горбунов А.Ю., Гладчук А.С., Калниня Я.К., Шиловских В.В., Томилин Н.В., Суходолов Н.Г., Радилов А.С., Подольская Е.П.Babakov V.N., Gorbunov A.Y., Gladchuk A.S., Kalninya Y.K., Shilovskikh V.V., Tomilin N.V., Sukhodolov N.G., Radilov A.S., Podolskaya E.P.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.extrememedicine.ru/jour/article/view/84

Аддукты белков крови с фосфорорганическими соединениями (ФОС) как маркеры интоксикации позволяют не только установить факт отравления, но и идентифицировать ФОС по присоединившемуся остатку. Целью работы было разработать методику специфичной и селективной экстракции аддуктов белков крови с ФОС на поверхности матрично-активированной лазерной десорбцией/ионизацией (МАЛДИ) мишени, функционализированной мультимолекулярными структурами на основе стеарата лантана, с помощью металл-аффинной хроматографии. Показано, что сорбент способен удерживать как полноразмерные, так и деалкилированные аддукты белков крови с ФОС. С помощью предложенной методики были экстрагированы и идентифицированы пептиды сывороточного альбумина и бутирилхолинэстеразы человека, модифицированные различными ФОС, после инкубации плазмы крови человека с ФОС в диапазоне концентраций 1 × 100 нг/мл, что позволяет использовать этот подход для идентификации ФОС при расследовании реальных случаев интоксикаций.

As markers of intoxication, adducts of blood proteins with organophosphorus compounds (OPs) allow establishing the fact of poisoning and, furthermore, enable identification of the OPs by the attached residue. This study aimed to develop a method of specific and selective extraction of blood protein adducts carrying OPs on the surface of a matrix-assisted laser desorption/ionization (MALDI) target functionalized with multimolecular structures based on lanthanum stearate using metal affinity chromatography. We have shown the ability of adsorbent to retain both full-size and dealkylated adducts of blood proteins with OPs. The developed method allowed extraction and identification of peptides of human serum albumin and human butyrylcholinesterase modified with various OPs (after incubation of human blood plasma with OPs in concentrations from 1 to 100 ng/mL), which makes this approach applicable for the purposes of OPs identification in the context of investigation of real cases of intoxication.

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The authors declare that there are no conflicts of interest present.