@article{chu_sripada_reese_bhandari_adams_sly_crapanzano_menegatti_2021, title={Purification of polyclonal immunoglobulin G from human serum using peptide-based adsorbents}, volume={10}, ISSN={["1547-5905"]}, DOI={10.1002/aic.17482}, abstractNote={AbstractThis study presents the chromatographic purification of immunoglobulin G (IgG) from human plasma using a two‐column process integrating the peptide‐based adsorbents LigaGuard™, which captures non‐Ig plasma proteins in flow‐through mode, and LigaTrap™, which isolates IgG in bind‐and‐elute. Buffer composition and column loading were optimized for both adsorbents. Two process configurations were evaluated. In the first design, plasma was fed to a LigaGuard™ column to capture plasma proteins, the effluent was loaded on the LigaTrap™ column, and the bound IgG was eluted with 63.8% global recovery and 99.7% purity; in comparison, Protein G agarose afforded approximately 67% recovery and 97.2% purity. In the alternative design, the LigaGuard™ column was utilized to polish the LigaTrap™ elution stream, affording 82.3% global recovery and 98.8% purity. Collectively, these results demonstrate the potential of a fully chromatographic process for purifying polyclonal IgG from plasma feedstocks.}, journal={AICHE JOURNAL}, author={Chu, Wenning and Sripada, Sobhana A. and Reese, Hannah R. and Bhandari, Dipendra and Adams, Augustus and Sly, Jae and Crapanzano, Michael and Menegatti, Stefano}, year={2021}, month={Oct} }
 @article{reese_bordelon_odeh_broussard_kormos_murphy_shanahan_menegatti_2020, title={Purification of animal immunoglobulin G (IgG) using peptoid affinity ligands}, ISBN={1520-6033}, DOI={10.1002/btpr.2994}, abstractNote={AbstractThe availability of highly pure animal antibodies is critical in the production of diagnostic tools and biosensors. The peptoid PL16, previously isolated from an ensemble of peptoid variants of the IgG‐binding peptide HWRGWV, was utilized in this work as affinity ligand on WorkBeads resin for the purification of immunoglobulin G (IgG) from a variety of mammalian sources and chicken immunoglobulin Y (IgY). The chromatographic protocol initially optimized for murine serum and ascites was subsequently employed for processing rabbit, goat and sheep, donkey, llama, and chicken sera. The PL16‐WorkBeads resin proved able to recover all antibody targets with values of yield between 50 and 90%, and purity consistently above 90%. Notably, PL16 not only binds a broader spectrum of animal immunoglobulins than the reference ligands Protein A and G, but it also binds equally well with all their subclasses. Unlike the protein ligands, in fact, PL16 afforded excellent values of yield and purity of mammalian polyclonal IgG, namely murine (47 and 94%), rabbit (66.5 and 91.7%), caprine IgG (63 and 91–95%), donkey, and llama (93 and 97%), as well as chicken IgY (42 and 92%). Of notice, it is also the ability of PL16 to target monomeric IgG without binding aggregated IgG; when challenged with a mixture of monomeric and aggregated murine IgG, PL16 eluted <3% of fed aggregates, against 11–13% eluted by Protein A and G. Collectively, these results prove the potential of the proposed peptoid ligand for large‐scale purification of animal immunoglobulins.}, journal={BIOTECHNOLOGY PROGRESS}, author={Reese, Hannah and Bordelon, Tee and Odeh, Fuad and Broussard, Amanda and Kormos, Chad and Murphy, Andrew and Shanahan, Calvin and Menegatti, Stefano}, year={2020} }
 @article{bacon_bowen_reese_rao_menegatti_2020, title={Use of Target-Displaying Magnetized Yeast in Screening mRNA-Display Peptide Libraries to Identify Ligands}, volume={22}, ISSN={["2156-8944"]}, DOI={10.1021/acscombsci.0c00171}, abstractNote={This work presents the first use of yeast-displayed protein targets for screening mRNA-display libraries of cyclic and linear peptides. The WW domains of Yes-Associated Protein 1 (WW-YAP) and mitochondrial import receptor subunit TOM22 were adopted as protein targets. Yeast cells displaying WW-YAP or TOM22 were magnetized with iron oxide nanoparticles to enable the isolation of target-binding mRNA-peptide fusions. Equilibrium adsorption studies were conducted to estimate the binding affinity (KD) of select WW-YAP-binding peptides: KD values of 37 and 4 μM were obtained for cyclo[M-AFRLC-K] and its linear cognate, and 40 and 3 μM for cyclo[M-LDFVNHRSRG-K] and its linear cognate, respectively. TOM22-binding peptide cyclo[M-PELNRAI-K] was conjugated to magnetic beads and incubated with yeast cells expressing TOM22 and luciferase. A luciferase-based assay showed a 4.5-fold higher binding of TOM22+ yeast compared to control cells. This work demonstrates that integrating mRNA- and yeast-display accelerates the discovery of peptide ligands.}, number={12}, journal={ACS COMBINATORIAL SCIENCE}, author={Bacon, Kaitlyn and Bowen, John and Reese, Hannah and Rao, Balaji M. and Menegatti, Stefano}, year={2020}, month={Dec}, pages={738–744} }