Comparison of TCA and collagenase in the isolation of tissue collagen

✍ Scribed by Robert A. Newman; Ronald O. Langner

• Publisher: Elsevier Science
• Year: 1975
• Tongue: English
• Weight: 631 KB
• Volume: 66
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(75)90735-6

✦ Synopsis

Studies on collagen metabolism often require the separation of collagen from other noncollagen proteins. The two major methods of isolating collagen are hot (90°C) trichloroacetic acid (TCA) extraction and/or collagenase solubilization. Though both procedures are widely used, a direct comparison and evaluation of their specificity and efficiency has not been investigated. In this study the specificity of both methods was examined using [Yltryptophan labeled proteins and the efficiency of collagen isolation was examined using a [Ylglycine labeled rat aortic homogenate. Protease free collagenase was highly specific in solubilizing collagen, while the hot TCA extract contained up to 14% of ['Kltryptophanlabeled noncollagen proteins. The hot TCA method consistently removed greater than 88% of total collagen from aortic tissue while the collagenase solubilization efficiency was less than 65%. Both procedures produced pure collagen fractions based on proline/hydroxyproline ratios. These data indicate that hot TCA yields a more complete extraction than does collagenase solubilization, but collagenase is' more specific in its solubilization of collagen tissue proteins.