@article{liu_gurgel_carbonell_2013, title={Affinity chromatographic purification of human immunoglobulin M from human B lymphocyte cell culture supernatant}, volume={70}, ISSN={["1873-295X"]}, DOI={10.1016/j.bej.2012.10.003}, abstractNote={Abstract Compared to immunoglobulin G purification with extensively studied affinity ligands such as protein A and protein G, little work has been done on affinity chromatographic purification of immunoglobulin M. Hexamer peptide ligand HWRGWV, previously shown to bind specifically to the Fc fragment of IgG, also demonstrated potential for IgM purification. This study presents further characterization and investigation of this ligand for its potential for purification of IgM. Different running conditions were employed in order to improve the recovery and purity of IgM. The final recovery and purity of the antibody is feedstock dependent, but can reach levels of both recovery and purity as high as 95%. The dependence of the recovery and purity on total loading amount and initial IgM concentration were investigated and discussed. Although relatively low dynamic binding capacities (DBC) in the range of 4.6–13.1 mg IgM/mL resin at linear flow rates from 173 to 35 cm/h were obtained for IgM compared to IgG because of the large molecular weight of IgM, the DBC value of HWRGWV for IgM is much greater than protein-based IgM affinity ligands found in the literature and is competitive with current commercially available affinity ligands, such as KAPTIVE-M, CaptureSelect IgM and Ultralink Immobilized Mannan Binding Protein.}, journal={BIOCHEMICAL ENGINEERING JOURNAL}, author={Liu, Zhuo and Gurgel, Patrick V. and Carbonell, Ruben G.}, year={2013}, month={Jan}, pages={63–70} }
 @article{liu_gurgel_carbonell_2013, title={Affinity chromatographic purification of human immunoglobulin a from chinese hamster ovary cell culture supernatant}, volume={29}, ISSN={["1520-6033"]}, DOI={10.1002/btpr.1652}, abstractNote={Abstract}, number={1}, journal={BIOTECHNOLOGY PROGRESS}, author={Liu, Zhuo and Gurgel, Patrick V. and Carbonell, Ruben G.}, year={2013}, pages={91–98} }
 @article{liu_gurgel_carbonell_2012, title={Purification of human immunoglobulins A, G and M from Cohn fraction II/III by small peptide affinity chromatography}, volume={1262}, ISSN={["1873-3778"]}, DOI={10.1016/j.chroma.2012.09.026}, abstractNote={This work describes attempts to purify human IgG, IgA and IgM from Cohn fraction II/III using HWRGWV affinity peptide resin. The effects of peptide density and different elution additives on recovery of the three antibodies were investigated. At low peptide density, salting-in salts such as magnesium chloride and calcium chloride facilitated antibody elution. Ethylene glycol, urea and arginine also facilitated elution because of their ability to decrease hydrophobic interactions, hydrogen bonding and electrostatic interactions. However, at high peptide density, no recovery improvements were observed because of increased non-specific hydrophobic interactions. The final elution conditions for each antibody were chosen based on the resulting yields and purities when a 10:2:1 mg/mL mixture of human IgG, IgA and IgM was used as starting material. Different pretreatment methods were employed in order to improve the purity of antibodies from Cohn fraction II/III. After pretreatment with caprylic acid precipitation or combination of caprylic acid and polyethylene glycol precipitation, purities over 95% and yields of about 60% were obtained for hIgG, which are comparable to current chromatographic purification methods involving two chromatography steps when hIgG is isolated from plasma fractions. A hIgA-enriched fraction with 42% hIgA and 56% hIgG, as well as a hIgM enriched fraction with 46% hIgM, 28% hIgA and 24% hIgG, were obtained as the by-products.}, journal={JOURNAL OF CHROMATOGRAPHY A}, author={Liu, Zhuo and Gurgel, Patrick V. and Carbonell, Ruben G.}, year={2012}, month={Nov}, pages={169–179} }
 @article{liu_gurgel_carbonell_2011, title={Effects of peptide density and elution pH on affinity chromatographic purification of human immunoglobulins A and M}, volume={1218}, ISSN={["1873-3778"]}, DOI={10.1016/j.chroma.2011.09.038}, abstractNote={A family of linear hexamer peptide ligands HWRGWV, HYFKFD and HFRRHL, initially identified for their affinity to the Fc portion of human immunoglobulin G (hIgG), also have potential for use in the purification of human immunoglobulins A (hIgA) and M (hIgM). HWRGWV demonstrated the strongest binding affinity to hIgM, followed by hIgA and hIgG respectively. The effects of N-terminal acetylation of the peptide, as well as elution buffer pH, on the chromatographic elution of human IgG, IgA and IgM from HWRGWV resins at various peptide densities (0.04–0.55 meq/g) were investigated. Over 80% recovery and 90% purity were achieved for human IgG and IgA isolation from complete minimum essential medium (cMEM) using HWRGWV resin at optimum peptide densities. For human IgM, 75.7% recovery and 86.0% purity were achieved by using HWRGWV at a low peptide density of 0.04 meq/g. Although HYFKFD and HFRRHL exhibited their ability for isolation of human IgG, IgA and IgM from cMEM as well, HWRGWV is the best option among them for large-scale purification of human IgG, IgA and IgM based on conditions tested.}, number={46}, journal={JOURNAL OF CHROMATOGRAPHY A}, author={Liu, Zhuo and Gurgel, Patrick V. and Carbonell, Ruben G.}, year={2011}, month={Nov}, pages={8344–8352} }