Simple and rapid chromatographic purification of type V collagen from a pepsin digest of porcine intestinal connective tissue, an unmanageable starting material for conventional column chromatography.
A chromatographic method is described for purification of type V collagen, a minor constituent in extracellular matrix, from a pepsin digest of porcine intestinal connective tissue. The starting material was a viscous and turbid solution even after centrifugation. Direct application of the sample to a commercially available DEAE-cellulose column resulted in clogging. On the other hand, type V collagen, [alpha1(V)](2)alpha2(V) form, was successfully captured by a filter paper-based DEAE-cellulose column chromatography and purified by a subsequent commercially available cation-exchange medium without clogging. This is a vast improvement over previously described salt fractionation methods.
Sato
Kenji
K
Department of Food Sciences and Nutritional Health, Kyoto Prefectural University, Shimogamao, 606-8522, Kyoto, Japan. k_sato@kpu.ac.jp
Tanahashi-Shiina
Tamae
T
Jun
Feng
F
Watanabe-Kawamura
Atsuko
A
Ichinomiya
Masami
M
Minegishi
Yutaka
Y
Tsukamasa
Yasuyuki
Y
Nakamura
Yasushi
Y
Kawabata
Makoto
M
Ohtsuki
Kozo
K
D000818:Animals; D002848:Chromatography, DEAE-Cellulose / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D024062:Collagen Type V / Q000302:isolation & purification*; D003238:Connective Tissue / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism*; D010434:Pepsin A / Q000378:metabolism*; D013552:Swine
D024062:Collagen Type V; D010434:Pepsin A