Establishment of an indirect ELISA for detection of the novel antifibrotic peptide M10.
Establishment of an indirect ELISA for detection of the novel antifibrotic peptide M10.
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DOI：
10.1371/journal.pone.0188588
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Bogatkevich GS
中科院分区：
文献类型：
--
作者：
Akter T;Atanelishvili I;Noguchi A;Silver RM;Bogatkevich GS
M10是由肝细胞生长因子（HGF）受体c-Met的胞内细胞质尾部被caspase-3切割后产生的10个氨基酸肽。最近，我们报道了M10与Smad2相互作用，并在体外和体内表现出抗纤维化特性，并可能成为一种新的抗纤维化药物。目前的研究是为了开发一种免疫分析法来测量生物标本中的M10浓度。建立了一种用于检测生物体液中M10的间接酶联免疫吸附试验（ELISA），使用制药级合成M10作为校准剂和市售的抗c- met C12抗体。小鼠单次腹腔注射M10 （1mg/kg）后，ELISA在血浆中特异性检测到M10，但不检测到混乱肽。检出限为9.6 ng/ml，测量限在15 ~ 200 ng/ml之间。M10的回收率在80% ~ 120%之间；试验内变异系数为5.3% ~ 6.3%；在15 ~ 250 ng /ml缓冲液浓度范围内，测定间变异系数在5.0% ~ 8.0%之间。单次腹腔注射（1mg/kg）后，M10浓度在6小时内达到峰值，并在48小时内降至最低水平。实验得到的M10半衰期与理论预测的M10半衰期相当。我们建立了一种高灵敏度的ELISA检测血浆样品中的抗纤维化肽M10，这将成为研究M10治疗纤维增生性疾病的药代动力学和疗效的新工具。
M10 is a ten amino acid peptide generated from the intracellular cytoplasmic tail of the hepatocyte growth factor (HGF) receptor c-Met following cleavage by caspase-3. Recently we reported that M10 interacts with Smad2 and demonstrates antifibrotic properties in vitro and in vivo and can be advanced into a novel antifibrotic remedy. The current study was undertaken to develop an immunoassay to measure M10 concentration in biological specimens. An Indirect Enzyme-Linked Immunosorbent Assay (ELISA) for detection of M10 in biological fluids was developed using pharmaceutical grade synthetic M10 as a calibrator and commercially available anti-c-Met C12 antibody. M10 ELISA specifically detected in plasma M10, but not a scrambled peptide, following a single intraperitoneal administration of M10 (1mg/kg) to mice. The detection limit was 9.6 ng/ml, and the measuring limit was between 15 ng/ml and 200 ng/ml. The recovery limits of M10 were between 80% and 120%; intra-assay coefficient of variation was between 5.3% and 6.3%; inter-assay coefficient of variation was between 5.0% and 8.0% over the buffer concentration tested in the range from 15 ng /ml to 250 ng /ml. The peak of M10 concentration following a single intraperitoneal injection (1mg/kg) was achieved within 6 hours and declined to minimal levels by 48 hours. The experimentally obtained half-life for M10 was comparable to the theoretically predicted half-life for M10. We have established a highly sensitive ELISA to detect the antifibrotic peptide M10 in plasma samples, which should prove to be a novel tool to study the pharmacokinetics and efficacy of M10 in the treatment of fibroproliferative disorders.
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影响因子：
13.5
作者：
Ma, Jihong;Zou, Chunbin;Guo, Lida;Seneviratne, Danushka S.;Tan, Xinping;Kwon, Yong-Kook;An, Jiyan;Bowser, Robert;DeFrances, Marie C.;Zarnegar, Reza
通讯作者：
Zarnegar, Reza
影响因子：
3.7
作者：
Atanelishvili, Ilia;Shirai, Yuichiro;Bogatkevich, Galina S.
通讯作者：
Bogatkevich, Galina S.
影响因子：
56.9
作者：
BACHMAIR, A;FINLEY, D;VARSHAVSKY, A
通讯作者：
VARSHAVSKY, A
DOI：
10.1016/j.trsl.2015.12.009
发表时间：
2016-04
期刊：
Translational research : the journal of laboratory and clinical medicine
影响因子：
--
作者：
Atanelishvili I;Shirai Y;Akter T;Buckner T;Noguchi A;Silver RM;Bogatkevich GS
通讯作者：
Bogatkevich GS
影响因子：
5.2
作者：
Neuss, S;Becher, E;Jahnen-Dechent, W
通讯作者：
Jahnen-Dechent, W