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 -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.
Issue Section:
Technical Briefs
1.
Kadler
, K. E.
, Holmes
, D. F.
, Trotter
, J. A.
, and Chapman
, J. A.
, 1996, “Collagen Fibril Formation
,” Biochem. J.
0264-6021, 316
, pp. 1
–11
.2.
Gross
, J.
, Highberger
, J. H.
, and Schmitt
, F. O.
, 1951, “A New Fibrous Structure Obtained from Extracts of Collagenous Connective Tissue
,” J. Appl. Phys.
0021-8979, 22
(1
), pp. 112
–112
.3.
Gross
, J.
, Highberger
, J. H.
, and Schmitt
, F. O.
, 1952, “Some Factors Involved in the Fibrogenesis of Collagen Invitro
,” in Proceedings of the Society for Experimental Biology and Medicine
, Vol. 80
(3
), pp. 462
–465
.4.
Gross
, J.
, Highberger
, J. H.
, and Schmitt
, F. O.
, 1954, “Collagen Structures Considered as States of Aggregation of a Kinetic Unit—the Tropocollagen Particle
,” in Proceedings of the National Academy of Sciences of the United States of America
, Vol. 40
(8
), pp. 679
–688
.5.
Gross
, J.
, 1956, “The Behaviour of Collagen Units as a Model in Morphogenesis
,” J. Biophys. Biochem. Cytol.
0095-9901, 25;2
(4
, Suppl.), pp. 261
–274
.6.
Gross
, J.
, and Kirk
, D.
, 1958, “Heat Precipitation of Collagen from Neutral Salt Solutions—Some Rate-Regulating Factors
,” J. Biol. Chem.
0021-9258, 233
(2
), pp. 355
–360
.7.
Wood
, G. C.
, and Keech
, M. K.
, 1960, “Formation of Fibrils from Collagen Solutions. 1. Effect of Experimental Conditions—Kinetic and Electron-Microscope Studies
,” Biochem. J.
0264-6021, 75
, pp. 588
–598
.8.
Wood
, G. C.
, 1960, “Formation of Fibrils from Collagen Solutions. 2. Mechanism of Collagen-Fibril Formation
,” Biochem. J.
0264-6021, 75
, pp. 598
–605
.9.
Bard
, J. B.
, and Chapman
, J. A.
, 1968, “Polymorphism in Collagen Fibrils Precipitated at Low pH
,” Nature (London)
0028-0836, 219
(160
), pp. 1279
–1280
.10.
Bard
, J. B. L.
, and Chapman
, J. A.
, 1973, “Diameters of Collagen Fibrils Grown In Vitro
,” Nature (London), New Biol.
0369-4887, 246
(151
), pp. 83
–84
.11.
Orekhovich
, V. N.
, Tustanovskii
, A. A.
, Orekhovich
, K. D.
, and Plotnikova
, N. E.
, 1948, “O Prokollagene Kozhi
,” Biochemistry (Mosc.)
0006-2979, 13
(1
), pp. 55
–60
.12.
Highberger
, J. H.
, Gross
, J.
, and Schmitt
, F. O.
, 1951, “The Interaction of Mucoprotein with Soluble Collagen—An Electron Microscope Study
,” Proc. Natl. Acad. Sci. U.S.A.
0027-8424, 37
(5
), pp. 286
–291
.13.
Binnig
, G.
, Quate
, C. F.
, and Gerber
, C.
, 1986, “Atomic Force Microscope
,” Phys. Rev. Lett.
0031-9007, 56
(9
), pp. 930
–933
.14.
Shattuck
, M. B.
, Gustafsson
, M. G. L.
, Fisher
, K. A.
, Yanagimoto
, K. C.
, Veis
, A.
, Bhatnagar
, R. S.
, and Clarke
, J.
, 1994, “Monomeric Collagen Imaged by Cryogenic Force Microscopy
,” J. Microsc.
0022-2720, 174
, pp. Rp1
–Rp2
.15.
Fujita
, Y.
, Kobayashi
, K.
, and Hoshino
, T.
, 1997, “Atomic Force Microscopy of Collagen Molecules. Surface Morphology of Segment-Long-Spacing (SLS) Crystallites of Collagen
,” J. Electron Microsc.
0022-0744, 46
(4
), pp. 321
–326
.16.
Paige
, M. F.
, Rainey
, J. K.
, and Goh
, M. C.
, 1998, “Fibrous Long Spacing Collagen Ultrastructure Elucidated by Atomic Force Microscopy
,” Biophys. J.
0006-3495, 74
(6
), pp. 3211
–3216
.17.
Morris
, V. J.
, Kirby
, A. R.
, and Gunning
, A. P.
, 1999, Atomic Force Microscopy for Biologists
, Imperial College Press
, London
.18.
Rainey
, J. K.
, Wen
, C. K.
, and Goh
, M. C.
, 2002, “Hierarchical Assembly and the Onset of Banding in Fibrous Long Spacing Collagen Revealed by Atomic Force Microscopy
,” Matrix Biol.
0945-053X, 21
(8
), pp. 647
–660
.19.
Gutsmann
, T.
, Fantner
, G. E.
, Venturoni
, M.
, Ekani-Nkodo
, A.
, Thompson
, J. B.
, Kindt
, J. H.
, Morse
, D. E.
, Fygenson
, D. K.
, and Hansma
, P. K.
, 2003, “Evidence that Collagen Fibrils in Tendons are Inhomogeneously Structured in a Tubelike Manner
,” Biophys. J.
0006-3495, 84
(4
), pp. 2593
–2598
.20.
Franzblau
, C.
, Schmid
, K.
, Faris
, B.
, Beldekas
, J.
, Garvin
, P.
, Kagan
, H. M.
, and Baum
, B. J.
, 1976, “Interaction of Collagen with Alpha-1-Acid Glycoprotein
,” Biochim. Biophys. Acta
0006-3002, 427
(1
), pp. 302
–314
.21.
Newman
, S.
, Cloitre
, M.
, Allain
, C.
, Forgacs
, G.
, and Beysens
, D.
, 1997, “Viscosity and Elasticity During Collagen Assembly In Vitro: Relevance to Matrix-Driven Translocation
,” Biopolymers
0006-3525, 41
(3
), pp. 337
–347
.22.
Pappenheimer
, J. R.
, Renkin
, E. M.
, and Borrero
, L. M.
, 1951, “Filtration, Diffusion and Molecular Sieving through Peripheral Capillary Membranes a Contribution to the Pore Theory of Capillary Permeability
,” Am. J. Physiol.
0002-9513, 167
(1
), pp. 13
–46
.23.
Pappenheimer
, J. R.
, 1953, “Passage of Molecules Through Capillary Walls
,” Physiol. Rev.
0031-9333, 33
(3
), pp. 387
–423
.24.
Renkin
, E. M.
, 1954, “Filtration, Diffusion, and Molecular Sieving through Porous Cellulose Membranes
,” J. Gen. Physiol.
0022-1295, 38
(2
), pp. 225
–243
.25.
Feins
, M.
, and Sirkar
, K. K.
, 2004, “Highly Selective Membranes in Protein Ultrafiltration
,” Biotechnol. Bioeng.
0006-3592, 86
(6
), pp. 603
–611
.26.
Kiesslich
, J.
, Radacher
, M.
, Neuhuber
, F.
, Meyer
, H. J.
, and Zeller
, K. W.
, 2002, “On the Use of Nitrocellulose Membranes for Dialysis-Mediated Purification of Ancient DNA from Human Bone and Teeth Extracts
,” Ancient Biomolecules
, 4
(2
), pp. 79
–87
.27.
Ruckenstein
, E.
, and Zeng
, X. F.
, 1997, “Macroporous Chitin Affinity Membranes for Lysozyme Separation
,” Biotechnol. Bioeng.
0006-3592, 56
(6
), pp. 610
–617
.28.
Atamanenko
, I.
, Kryvoruchko
, A.
, and Yurlova
, L.
, 2004, “Study of the Scaling Process on Membranes
,” Desalination
0011-9164, 167
(1–3
), pp. 327
–334
.29.
Bosch
, R.
, Lautenschlager
, P.
, Potthast
, L.
, and Stapelmann
, J.
, 1992, “Experiment Equipment for Protein Crystallization in Mu-G Facilities
,” J. Cryst. Growth
0022-0248, 122
(1–4
), pp. 310
–316
.30.
Vergara
, A.
, Corvino
, E.
, Sorrentino
, G.
, Piccolo
, C.
, Tortora
, A.
, Carotenuto
, L.
, Mazzarella
, L.
, and Zagari
, A.
, 2002, “Crystallization of the Collagen-Like Polypeptide (PPG)(10) Aboard the International Space Station. 1. Video Observation
,” Acta Crystallogr., Sect. D: Biol. Crystallogr.
0907-4449, 58
, pp. 1690
–1694
.31.
Berisio
, R.
, Vitagliano
, L.
, Sorrentino
, G.
, Carotenuto
, L.
, Piccolo
, C.
, Mazzarella
, L.
, and Zagari
, A.
, 2000, “Effects of Microgravity on the Crystal Quality of a Collagen-Like Polypeptide
,” Acta Crystallogr., Sect. D: Biol. Crystallogr.
0907-4449, 56
, pp. 55
–61
.32.
Graham
, J. S.
, Vomund
, A. N.
, Phillips
, C. L.
, and Grandbois
, M.
, 2004, “Structural Changes in Human Type I Collagen Fibrils Investigated by Force Spectroscopy
,” Exp. Cell Res.
0014-4827, 299
(2
), pp. 335
–342
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