Comparison of five procedures for the purification of insoluble elastin

Elastin is an insoluble, highly cross-linked protein, providing elasticity to organs like lung, aorta, and ligaments. Despite its remarkable mechanical properties, elastin has found little use as a biomaterial. Purification of intact elastin from elastic fibres presents a major challenge, among othe...

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Bibliographic Details
Published in:Biomaterials 2001-07, Vol.22 (14), p.1997-2005
Main Authors: Daamen, W.F, Hafmans, T, Veerkamp, J.H, van Kuppevelt, T.H
Format: Article
Language:English
Subjects:
Acetone - pharmacology
Amino Acids - analysis
Animals
Biological and medical sciences
Calcification
Chemical Fractionation - methods
Collagenases - pharmacology
Cyanogen Bromide - pharmacology
Elastin - chemistry
Elastin - immunology
Elastin - isolation & purification
Guanidine - pharmacology
Horses
Hot Temperature
Insoluble elastin
Ligaments - chemistry
Ligaments - drug effects
Ligamentum nuchae
Medical sciences
Mercaptoethanol - pharmacology
Microscopy, Electron
Microscopy, Fluorescence
Purification
Sodium Hydroxide - pharmacology
Solubility
Solvents - pharmacology
Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases
Technology. Biomaterials. Equipments
Trypsin - pharmacology
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Summary:Elastin is an insoluble, highly cross-linked protein, providing elasticity to organs like lung, aorta, and ligaments. Despite its remarkable mechanical properties, elastin has found little use as a biomaterial. Purification of intact elastin from elastic fibres presents a major challenge, among others for the intimate interwoveness of elastin and microfibrils. Insoluble elastin preparations tend to calcify, which may be due to calcium-binding microfibrillar (e.g. fibrillin). In this study, elastin was purified from horse ligamentum nuchae using five different procedures. One procedure is based on treatment with 0.1 m NaOH, another on autoclaving and treatment with cyanogen bromide. Three other procedures are based on combinations of extraction steps and enzyme digestions. Purity of preparations was assessed by sodium dodecyl sulphate polyacrylamide gel electrophoresis, amino acid analysis, bright field immunofluorescence and transmission electron microscopy. The procedure involving extractions/enzymes combined with an early application of 2-mercaptoethanol and cyanogen bromide gives a highly pure elastin preparation. Electron microscopic analysis showed that this preparation is devoid of microfibrillar components. This procedure is therefore the method of choice for preparation of insoluble elastin as a biomaterial for tissue engineering.
ISSN:0142-9612
1878-5905
DOI:10.1016/S0142-9612(00)00383-5