title: Heat elution chromatography of immunoglobulins.
abstract: A tremendous increase has taken place over the last decades in the biochemical and clinical use of antibodies. Unfortunately, the constantly growing demand has not been matched by a corresponding easy access to pure immunoglobulin, as purification procedures tend to be either laborious, expensive, or inefficient. We present a new and simplified method to obtain pure antibody based on the special thermal properties of the streptococcal M proteins, a family of cell-surface exposed coiled-coil molecules which bind different sets of host plasma proteins. The coiled-coil structure is already destabilized at low temperatures and the M proteins unfold reversibly, usually below 40 degrees C. We demonstrate the use of this property to purify immunoglobulin G from rabbit serum with protein H from the AP1 strain of Streptococcus pyogenes. Recombinant protein H is linked to nickel-agarose via a C-terminal histidine tag. After mixing with rabbit serum and washing at room temperature, pure IgG can be eluted from the gel with a moderately heated buffer. In this case, protein H has been used to purify rabbit IgG, but the principle should be applicable to other M protein-ligand pairs.
authors.affiliation: Department of Cell and Molecular Biology, BMC, Plan C11, Lund University, SE-22184 Lund, Sweden. peter.osmark@medkem.lu.se
authors.forename: Peter
authors.initials: P
authors.lastname: Osmark
authors.forename: Tommy
authors.initials: T
authors.lastname: Cedervall
authors.forename: Katrien
authors.initials: K
authors.lastname: Pieters
authors.forename: Bo
authors.initials: B
authors.lastname: Akerström
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000378:metabolism; D002845:Chromatography / Q000379:methods*; D006358:Hot Temperature; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D011817:Rabbits
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D007074:Immunoglobulin G; D008565:Membrane Proteins; C064006:protein H, Streptococcus