Validation of Therapeutic Agent Conjugation to Polyvinyl Alcohol-Coated Medical Devices.

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES
Joshua C Colvin, Donald Sorrells, J Steven Alexander
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引用次数: 0

Abstract

Protein-based therapeutics are often limited by their route of administration and inability to confine them to their site of action. One innovative approach we have developed is to covalently bind protein therapeutics to medical devices, allowing more localized and highly concentrated delivery of these agents to their intended site of action. This study aims to evaluate if glucagon-like peptide-2 (GLP-2) can be covalently bound to the vaginal expansion sleeve (VES) and intestinal expansion sleeve (IES) devices in clinically relevant and measurable quantities. Expansion sleeves were coated with polyvinyl alcohol (PVA) and crosslinked with glutaraldehyde/sulfuric acid vapor to create a chemically active surface capable of binding amine-containing therapeutics such as GLP-2. A standard curve was created by binding 250 µg, 100 µg, 50 µg, 25 µg, and 0 µg of GLP-2 to PVA-coated wells of a 24-well plate. An ELISA standard was created using a rabbit anti-GLP-2 antibody followed by a goat anti-rabbit IgG alkaline phosphatase secondary antibody plus an alkaline phosphatase blue microwell substrate. Colorimetry (yellow to blue at 620 nm) was proportional to the concentration of GLP-2 bound antibodies, enabling calculation of the bound concentration of GLP-2 on the PVA-coated sleeves. The addition of 50 µg of GLP-2 to IES/VES devices bound an average of 22.69 ± 9.32 µg/cm2 of GLP-2 with an external IES/VES surface area (9.425 cm2), indicating that 44% of added GLP-2 was immobilized on the PVA coated IES/VES sleeves. Current human GLP-2 dosing is 50 µg/Kg. Because each sleeve carries 22 µg, this is approximately 44% of a systemic dose in a single device. This methodology makes it possible to add dramatically lower doses of therapeutic agents to get the same effect as systemic administration of the GLP-2 drug while also avoiding systemic effects.

以蛋白质为基础的疗法往往受到给药途径的限制,无法将其限制在作用部位。我们开发的一种创新方法是将蛋白质疗法与医疗设备共价结合,从而将这些药物更局部、更高浓度地输送到预定的作用部位。本研究旨在评估胰高血糖素样肽-2(GLP-2)能否共价结合到阴道扩张套(VES)和肠道扩张套(IES)装置上,并达到临床相关的可测量数量。膨胀套表面涂有聚乙烯醇 (PVA),并与戊二醛/硫酸蒸气交联,形成一个化学活性表面,能够与 GLP-2 等含胺治疗药物结合。将 250 微克、100 微克、50 微克、25 微克和 0 微克的 GLP-2 与涂有 PVA 的 24 孔板孔结合,即可绘制出标准曲线。使用兔抗 GLP-2 抗体、山羊抗兔 IgG 碱性磷酸酶二抗和碱性磷酸酶蓝色微孔底物建立 ELISA 标准。比色法(620 纳米波长处由黄转蓝)与 GLP-2 结合抗体的浓度成正比,从而计算出 PVA 涂层套管上 GLP-2 的结合浓度。在 IES/VES 装置中添加 50 µg GLP-2 后,IES/VES 的外部表面积(9.425 cm2)平均结合了 22.69 ± 9.32 µg/cm2 的 GLP-2,这表明添加的 GLP-2 有 44% 固定在涂有 PVA 的 IES/VES 套管上。目前人类 GLP-2 的剂量为 50 µg/Kg。由于每个套管可承载 22 µg,因此单个装置的剂量约为全身剂量的 44%。通过这种方法,可以大大降低治疗剂的剂量,从而获得与全身服用 GLP-2 药物相同的效果,同时还能避免全身效应。
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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
期刊介绍: JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.
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