Construction, expression, and purification of recombinant αVβ5 integrin.

Authors :: Tartaglia LJ
Bennett A
Woodhouse AG
Aydemir F
Muzyczka N
Agbandje-McKenna M
Source :: Protein expression and purification [Protein Expr Purif] 2013 Jun; Vol. 89 (2), pp. 225-31. Date of Electronic Publication: 2013 Apr 12.
Publication Year :: 2013
Abstract: A recombinant integrin expression system has been created for the large-scale production of αVβ5 integrin extracellular domains that take advantage of Fos and Jun dimerization for expression in bacterial, insect, and mammalian cells. This utilizes an all-in-one vector, pQE-TriSystem, with molecular machinery for parallel expression without the need of additional subcloning. Optimal expression in HEK293 cells was determined by a time course analysis. The heterodimer was purified in a one-step nickel column purification scheme, and the sequence and functional state were confirmed by mass spectrometry and inhibition assays, respectively. The yields of αVβ5 integrin obtained are in quantities suitable for multiple applications including structural biology and functional assays.<br /> (Copyright © 2013 Elsevier Inc. All rights reserved.)