title: Excessive Labeling Technique Provides a Highly Sensitive Fluorescent Probe for Real-time Monitoring of Biodegradation of Biopolymer Pharmaceuticals in vivo.
abstract: Recombinant proteins represent a large sector of the biopharma market. Determination of the main elimination pathways raises the opportunities to significantly increase their half-lives in vivo. However, evaluation of biodegradation of pharmaceutical biopolymers performed in the course of pre-clinical studies is frequently complicated. Noninvasive pharmacokinetic and biodistribution studies in living organism are possible using proteins conjugated with near-infrared dyes. In the present study we designed a highly efficient probe based on fluorescent dye self-quenching for monitoring of in vivo biodegradation of recombinant human butyrylcholinesterase. The maximum enhancement of integral fluorescence in response to degradation of an intravenously administered enzyme was observed 6 h after injection. Importantly, excessive butyrylcholinesterase labeling with fluorescent dye results in significant changes in the pharmacokinetic properties of the obtained conjugate. This fact must be taken into consideration during future pharmacokinetic studies using in vivo bioimaging.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, GSP-7, Moscow, Russia, 117997.
authors.forename: S S
authors.initials: SS
authors.lastname: Terekhov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, GSP-7, Moscow, Russia, 117997 ; Kazan Federal University, Kremlevskaya str. 18, 420008, Kazan, Republic of Tatarstan, Russia.
authors.forename: I V
authors.initials: IV
authors.lastname: Smirnov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, GSP-7, Moscow, Russia, 117997.
authors.forename: O G
authors.initials: OG
authors.lastname: Shamborant
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev Ave., 8, 630090, Novosibirsk, Russia.
authors.forename: M A
authors.initials: MA
authors.lastname: Zenkova
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev Ave., 8, 630090, Novosibirsk, Russia.
authors.forename: E L
authors.initials: EL
authors.lastname: Chernolovskaya
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev Ave., 8, 630090, Novosibirsk, Russia.
authors.forename: D V
authors.initials: DV
authors.lastname: Gladkikh
authors.affiliation: Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, 142290, Moscow region, Russia.
authors.forename: A N
authors.initials: AN
authors.lastname: Murashev
authors.affiliation: Branch of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Pushchino, 142290, Moscow region, Russia.
authors.forename: I A
authors.initials: IA
authors.lastname: Dyachenko
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, GSP-7, Moscow, Russia, 117997.
authors.forename: V D
authors.initials: VD
authors.lastname: Knorre
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, GSP-7, Moscow, Russia, 117997 ; Kazan Federal University, Kremlevskaya str. 18, 420008, Kazan, Republic of Tatarstan, Russia ; Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, 119334, Moscow, Russia.
authors.forename: A A
authors.initials: AA
authors.lastname: Belogurov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, GSP-7, Moscow, Russia, 117997.
authors.forename: N A
authors.initials: NA
authors.lastname: Ponomarenko
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, GSP-7, Moscow, Russia, 117997.
authors.forename: S M
authors.initials: SM
authors.lastname: Deyev
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Lavrentiev Ave., 8, 630090, Novosibirsk, Russia.
authors.forename: V V
authors.initials: VV
authors.lastname: Vlasov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, GSP-7, Moscow, Russia, 117997 ; Kazan Federal University, Kremlevskaya str. 18, 420008, Kazan, Republic of Tatarstan, Russia ; Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, 119334, Moscow, Russia.
authors.forename: A G
authors.initials: AG
authors.lastname: Gabibov
keywords: biodegradation; butyrylcholinesterase; fluorescent probe; in vivo bioimaging; pharmacokinetics; proteolysis; self-quenching