Synthesis and evaluation of an anti-MLC1 × anti-CD90 bispecific antibody for targeting and retaining bone-marrow-derived multipotent stromal cells in infarcted myocardium.
Synthesis and evaluation of an anti-MLC1 × anti-CD90 bispecific antibody for targeting and retaining bone-marrow-derived multipotent stromal cells in infarcted myocardium.
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在梗塞心肌中靶向和固定骨骼衍生的多能细胞的抗MLC1×抗CD90双特异性抗体的合成和评估。
DOI：
10.1021/bc200309h
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发表时间：
2011-08-17
影响因子：
4.7
通讯作者：
Woodside, Darren G.
中科院分区：
文献类型：
--
作者：
Gundlach, C. William;Caivano, Amy;Cabreira-Hansen, Maria da Graca;Gahremanpour, Amir;Brown, Wells S.;Zheng, Yi;McIntyre, Bradley W.;Willerson, James T.;Dixon, Richard A. F.;Perin, Emerson C.;Woodside, Darren G.
干细胞在心血管再生医学中应用的一个关键问题是它们在靶组织中的滞留。在这里，我们已经产生并评估了一种双特异性抗体异二聚体，旨在改善骨髓来源的多潜能基质细胞(BMMSC)在心肌梗死损伤的心脏组织中的保留。杂二聚体由抗人CD90单抗(克隆5E10)和抗肌球蛋白轻链1(克隆MLM508)单抗(克隆MLM508)组成。我们用聚乙二醇化的4-甲酰基苯甲酰胺部分修饰抗CD90抗体，摩尔取代比(MSR)为2.6，用6-肼烟酰胺部分修饰抗MLC1抗体的摩尔取代比(MSR)为0.9。共价修饰对mAb表位结合无明显不良影响。此外，抗CD90抗体与骨髓间充质干细胞的结合并不能阻止其分化为脂肪、软骨或成骨细胞。修饰的抗体在温和的条件下(RT，pH 6，1h)在催化剂(苯胺)的存在下结合，以允许快速生成共价双芳基肼，这是在A354监测到的。我们评估了构建物中每一单抗的表位免疫反应性。流式细胞仪检测结果表明，该双功能载体可与游离抗CD90单抗竞争的BMMSCs结合，证实修饰和交联对抗CD90互补决定区无不良影响。同样，基于ELISA法的检测表明，双特异性抗体与塑料固定的重组MLC1结合。过量的抗MLC1mAb竞争双特异性抗体结合。最后，平行板流动室实验表明，抗CD90×抗MLC1双特异性抗体构建体可诱导骨髓间充质干细胞与塑料固定的抗剪切力的MLC1发生黏附。我们使用的单抗既能结合人类抗原，又能结合相应的猪同系物。因此，抗CD90×抗MLC1双特异性抗体可用于急性心肌梗死的大型动物研究，并可为临床研究提供一个起点。
A key issue regarding the use of stem cells in cardiovascular regenerative medicine is their retention in target tissues. Here, we have generated and assessed a bispecific antibody heterodimer designed to improve the retention of bone marrow–derived multipotent stromal cells (BMMSC) in cardiac tissue damaged by myocardial infarction. The heterodimer comprises an anti-human CD90 monoclonal antibody (mAb) (clone 5E10) and an anti-myosin light chain 1 (MLC1) mAb (clone MLM508) covalently cross-linked by a bis-aryl hydrazone. We modified the anti-CD90 antibody with a pegylated-4-formylbenzamide moiety to a molar substitution ratio (MSR) of 2.6 and the anti-MLC1 antibody with a 6-hydrazinonicotinamide moiety to a MSR of 0.9. The covalent modifications had no significant deleterious effect on mAb epitope binding. Furthermore, the binding of anti-CD90 antibody to BMMSCs did not prevent their differentiation into adipo-, chondro-, or osteogenic lineages. Modified antibodies were combined under mild conditions (RT, pH 6, 1 h) in the presence of a catalyst (aniline) to allow for rapid generation of the covalent bis-aryl hydrazone, which was monitored at A354. We evaluated epitope immunoreactivity for each mAb in the construct. Flow cytometry demonstrated binding of the bispecific construct to BMMSCs that was competed by free anti-CD90 mAb, verifying that modification and cross-linking were not detrimental to the anti-CD90 complementarity-determining region. Similarly, ELISA-based assays demonstrated bispecific antibody binding to plastic-immobilized recombinant MLC1. Excess anti-MLC1 mAb competed for bispecific antibody binding. Finally, the anti-CD90 × anti-MLC1 bispecific antibody construct induced BMMSC adhesion to plastic-immobilized MLC1 that was resistant to shear stress, as measured in parallel-plate flow chamber assays. We used mAbs that bind both human antigens and the respective pig homologues. Thus, the anti-CD90 × anti-MLC1 bispecific antibody may be used in large animal studies of acute myocardial infarction and may provide a starting point for clinical studies.
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DOI：
10.1084/jem.172.1.363
发表时间：
1990-07-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Lansdorp PM;Sutherland HJ;Eaves CJ
通讯作者：
Eaves CJ
DOI：
10.1016/0735-1097(88)91521-5
发表时间：
1988-03-01
影响因子：
24
作者：
KATUS, HA;DIEDERICH, KW;KUBLER, W
通讯作者：
KUBLER, W
影响因子：
37.8
作者：
KHAW, BA;GOLD, HK;HABER, E
通讯作者：
HABER, E
影响因子：
15
作者：
Dirksen, Anouk;Dirksen, Sjoerd;Dawson, Philip E.
通讯作者：
Dawson, Philip E.
影响因子：
24
作者：
Lipinski, Michael J.;Biondi-Zoccai, Giuseppe G. L.;Vetrovec, George W.
通讯作者：
Vetrovec, George W.