Strasser Stefan, Zink Albert, Heckl Wolfgang M, Thalhammer Stefan
Department für Geo- und Umweltwissenschaften, Ludwig-Maximilians-Universität, 80333 Munich, Germany.
J Biomech Eng. 2006 Oct;128(5):792-6. doi: 10.1115/1.2264392.
In vitro self-assembled collagen fibrils form a variety of different structures during dialysis. The self-assembly is dependent on several parameters, such as concentrations of collagen and alpha1-acid glycoprotein, temperature, dialysis time, and the acid concentration. For a detailed understanding of the assembly pathway and structural features like banding pattern or mechanical properties it is necessary to study single collagen fibrils. In this work we present a fully automated system to control the permeation of molecules through a membrane like a dialysis tubing. This allows us to ramp arbitrary diffusion rate profiles during the self-assembly process of macromolecules, such as collagen. The system combines a molecular sieving method with a computer assisted control system for measuring process variables. With the regulation of the diffusion rate it is possible to control and manipulate the collagen self-assembly process during the whole process time. Its performance is demonstrated by the preparation of various collagen type I fibrils and native collagen type II fibrils. The combination with the atomic force microscope (AFM) allows a high resolution characterization of the self-assembled fibrils. In principle, the represented system can be also applied for the production of other biomolecules, where a dialysis enhanced self-assembly process is used.
体外自组装的胶原纤维在透析过程中形成多种不同结构。自组装取决于几个参数，如胶原蛋白和α1-酸性糖蛋白的浓度、温度、透析时间以及酸浓度。为了详细了解组装途径和诸如条纹图案或机械性能等结构特征，有必要研究单个胶原纤维。在这项工作中，我们展示了一个全自动系统，用于控制分子透过类似透析管的膜的渗透。这使我们能够在大分子（如胶原蛋白）的自组装过程中设置任意的扩散速率曲线。该系统将分子筛分方法与用于测量过程变量的计算机辅助控制系统相结合。通过调节扩散速率，可以在整个过程时间内控制和操纵胶原蛋白的自组装过程。通过制备各种I型胶原纤维和天然II型胶原纤维证明了其性能。与原子力显微镜（AFM）结合可对自组装纤维进行高分辨率表征。原则上，所展示的系统也可应用于其他生物分子的生产，其中使用了透析增强的自组装过程。
