Heat elution chromatography of immunoglobulins Export

@article{citeulike:6016060, 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 °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.}, author = {Osmark, P.}, citeulike-article-id = {6016060}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/S1046-5928(03)00130-X}, citeulike-linkout-1 = {http://linkinghub.elsevier.com/retrieve/pii/S1046-5928(03)00130-X}, doi = {10.1016/S1046-5928(03)00130-X}, issn = {10465928}, journal = {Protein Expression and Purification}, keywords = {bag, bio, biochemical, bioreactor, case, cell, culture, engineering, igg, mab, process, processing, protocols, study, wave}, month = {August}, number = {2}, pages = {301--303}, posted-at = {2009-10-27 15:21:00}, priority = {4}, title = {Heat elution chromatography of immunoglobulins}, url = {http://dx.doi.org/10.1016/S1046-5928(03)00130-X}, volume = {30}, year = {2003} }

TY - JOUR ID - citeulike:6016060 L3 - citeulike-article-id:6016060 N2 - 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 °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. IS - 2 JF - Protein Expression and Purification SN - 10465928 EP - 303 TI - Heat elution chromatography of immunoglobulins VL - 30 SP - 301 KW - bag KW - bio KW - biochemical KW - bioreactor KW - case KW - cell KW - culture KW - engineering KW - igg KW - mab KW - process KW - processing KW - protocols KW - study KW - wave AU - Osmark, P PY - 2003/08// UR - http://dx.doi.org/10.1016/S1046-5928(03)00130-X ER -