Controlled self-assembly of collagen fibrils by an automated dialysis system.
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.
Strasser
Stefan
S
Department für Geo- und Umweltwissenschaften, Ludwig-Maximilians-Universität, 80333 Munich, Germany.
Zink
Albert
A
Heckl
Wolfgang M
WM
Thalhammer
Stefan
S
D003460:Crystallization / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D024022:Fibrillar Collagens / Q000737:chemistry* / Q000648:ultrastructure*; D017551:Microdialysis / Q000295:instrumentation* / Q000379:methods; D046912:Multiprotein Complexes / Q000737:chemistry / Q000648:ultrastructure
D024022:Fibrillar Collagens; D046912:Multiprotein Complexes