@article{sripada_barbieri_shastry_wuestenhagen_aldinger_rammo_schulte_daniele_menegatti_2024, title={Multiangle Light Scattering as a Lentivirus Purification Process Analytical Technology}, volume={5}, ISSN={["1520-6882"]}, DOI={10.1021/acs.analchem.4c01209}, abstractNote={The limited biomolecular and functional stability of lentiviral vectors (LVVs) for cell therapy poses the need for analytical tools that can monitor their titers and activity throughout the various steps of expression and purification. In this study, we describe a rapid (25 min) and reproducible (coefficient of variance ∼0.5–2%) method that leverages size exclusion chromatography coupled with multiangle light scattering detection (SEC-MALS) to determine size, purity, and particle count of LVVs purified from bioreactor harvests. The SEC-MALS data were corroborated by orthogonal methods, namely, dynamic light scattering (DLS) and transmission electron microscopy. The method was also evaluated for robustness in the range of 2.78 × 105–2.67 × 107 particles per sample. Notably, MALS-based particle counts correlated with the titer of infectious LVVs measured via transduction assays (R2 = 0.77). Using a combination of SEC-MALS and DLS, we discerned the effects of purification parameters on LVV quality, such as the separation between heterogeneous LV, which can facilitate critical decision-making in the biomanufacturing of gene and cell therapies.}, journal={ANALYTICAL CHEMISTRY}, author={Sripada, Sobhana A. and Barbieri, Eduardo and Shastry, Shriarjun and Wuestenhagen, Elena and Aldinger, Annika and Rammo, Oliver and Schulte, Michael M. and Daniele, Michael and Menegatti, Stefano}, year={2024}, month={May} }
 @article{barbieri_mollica_moore_sripada_shastry_kilgore_loudermilk_whitacre_kilgour_wuestenhagen_et al._2024, title={Peptide ligands targeting the vesicular stomatitis virus G (VSV-G) protein for the affinity purification of lentivirus particles}, volume={121}, ISSN={["1097-0290"]}, DOI={10.1002/bit.28594}, abstractNote={Abstract}, number={2}, journal={BIOTECHNOLOGY AND BIOENGINEERING}, author={Barbieri, Eduardo and Mollica, Gina N. and Moore, Brandyn D. and Sripada, Sobhana A. and Shastry, Shriarjun and Kilgore, Ryan E. and Loudermilk, Casee M. and Whitacre, Zachary H. and Kilgour, Katie M. and Wuestenhagen, Elena and et al.}, year={2024}, month={Feb}, pages={618–639} }
 @article{oh_mendola_choe_min_lavoie_sripada_williams_lee_yigzaw_seay_et al._2023, title={Identification and characterization of CHO host-cell proteins in monoclonal antibody bioprocessing}, volume={10}, ISSN={["1097-0290"]}, DOI={10.1002/bit.28568}, abstractNote={Abstract}, journal={BIOTECHNOLOGY AND BIOENGINEERING}, author={Oh, Young Hoon and Mendola, Kerri M. and Choe, Leila H. and Min, Lie and Lavoie, Ashton R. and Sripada, Sobhana A. and Williams, Taufika Islam and Lee, Kelvin H. and Yigzaw, Yinges and Seay, Alexander and et al.}, year={2023}, month={Oct} }
 @article{sripada_elhanafi_collins_williams_linova_woodley_boi_menegatti_2023, title={Pseudo-affinity capture of <i>K. phaffii</i> host cell proteins in flow-through mode: Purification of protein therapeutics and proteomic study}, volume={326}, ISSN={["1873-3794"]}, url={http://www.scopus.com/inward/record.url?eid=2-s2.0-85169830823&partnerID=MN8TOARS}, DOI={10.1016/j.seppur.2023.124777}, abstractNote={K. phaffii is a versatile expression system that is increasingly utilized to produce biological therapeutics – including enzymes, engineered antibodies, and gene-editing tools – that feature multiple subunits and complex post-translational modifications. Two major roadblocks limit the adoption of K. phaffii in industrial biomanufacturing: its proteome, while known, has not been linked to downstream process operations and detailed knowledge is missing on problematic host cell proteins (HCPs) that endanger patient safety or product stability. Furthermore, the purification toolbox has not evolved beyond the capture of monospecific antibodies, and few solutions are available for engineered antibody fragments and other protein therapeutics. To unlock the potential of yeast-based biopharmaceutical manufacturing, this study presents the development and performance validation of a novel adsorbent – PichiaGuard – functionalized with peptide ligands that target the whole spectrum of K. phaffii HCPs and designed for protein purification in flow-through mode. The PichiaGuard adsorbent features high HCP binding capacity (∼25 g per liter of resin) and successfully purified a monoclonal antibody and an ScFv fragment from clarified K. phaffii harvests, affording > 300-fold removal of HCPs and high product yields (70–80%). Notably, PichiaGuard outperformed commercial ion exchange and mixed-mode resins without salt gradients or optimization in removing high-risk HCPs – including aspartic proteases, ribosomal subunits, and other peptidases – thus demonstrating its value in modern biopharmaceutical processing.}, journal={SEPARATION AND PURIFICATION TECHNOLOGY}, author={Sripada, Sobhana A. and Elhanafi, Driss and Collins, Leonard B. and Williams, Taufika I. and Linova, Marina Y. and Woodley, John M. and Boi, Cristiana and Menegatti, Stefano}, year={2023}, month={Dec} }
 @article{fan_sripada_pham_linova_woodley_menegatti_boi_carbonell_2023, title={Purification of a monoclonal antibody using a novel high-capacity multimodal cation exchange nonwoven membrane}, volume={317}, ISSN={["1873-3794"]}, DOI={10.1016/j.seppur.2023.123920}, abstractNote={A high-capacity, multimodal cation exchange (MMC) chromatographic membrane was developed by conjugating a multimodal ligand – 2-mercaptopyridine-3-carboxylic acid (MPCA) – on a polybutylene terepthalate (PBT) nonwoven fabric. The membrane features an equilibrium binding capacity of ≈ 1000 mg of human polyclonal IgG (IgG) per g of membrane and dynamic binding capacities (DBC10%) ranging from 77.5 to 115.1 mg/mL (residence times of 1 and 5 min, respectively); these values are 2-to-3-fold higher than those of commercial MMC adsorbents. The effects of buffer composition, pH, conductivity on the binding behavior of the MMC-MPCA membrane were investigated in detail. As a moderate cation exchange binder, MPCA enables effective protein elution using buffers with mild pH (8.0–9.0) and conductivity (≈13 mS/cm), thus circumventing the harsh conditions often needed in multimodal chromatography. The MMC-MPCA membrane was evaluated for product capture in bind-and-elute mode on a Chinese hamster ovary (CHO) cell culture harvest containing therapeutic monoclonal antibodies, using commercial multimodal (Capto MMC and MX-Trp-650M) and affinity (AF-rProtein A HC-650F) resins as controls. The MMC-MPCA membrane outperformed the multimodal resins in terms of binding capacity as well as clearance of host cell proteins (HCPs) and aggregates. The membrane was then evaluated by polishing the mAb from a Protein A eluate in bind-and-elute mode. The MMC-MPCA membrane reduced the level of high molecular weight components from 11% to 4% and the HCP content from 1319.7 ppm to 48.7 ppm (LRV of 1.4). Most notably, proteomics analysis of the product demonstrated the clearance of a significant fraction of persistent, high-risk HCPs from the Protein A eluate.}, journal={SEPARATION AND PURIFICATION TECHNOLOGY}, author={Fan, Jinxin and Sripada, Sobhana A. and Pham, Dan N. and Linova, Marina Y. and Woodley, John M. and Menegatti, Stefano and Boi, Cristiana and Carbonell, Ruben G.}, year={2023}, month={Jul} }
 @article{sripada_chu_williams_teten_mosley_carbonell_lenhoff_cramer_bill_yigzaw_et al._2022, title={Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via "flow-through affinity chromatography" using peptide-based adsorbents}, volume={119}, ISSN={["1097-0290"]}, url={https://doi.org/10.1002/bit.28096}, DOI={10.1002/bit.28096}, abstractNote={Abstract}, number={7}, journal={BIOTECHNOLOGY AND BIOENGINEERING}, publisher={Wiley}, author={Sripada, Sobhana Alekhya and Chu, Wenning and Williams, Taufika Islam and Teten, Matthew A. and Mosley, Brian J. and Carbonell, Ruben G. and Lenhoff, Abraham M. and Cramer, Steven M. and Bill, Jerome and Yigzaw, Yinges and et al.}, year={2022}, month={Apr} }
 @article{sternisha_mukherjee_alex_chaney_barkalifa_wan_lee_rico-jimenez_zurauskas_spillman_et al._2021, title={Longitudinal monitoring of cell metabolism in biopharmaceutical production using label-free fluorescence lifetime imaging microscopy}, ISSN={["1860-7314"]}, DOI={10.1002/biot.202000629}, abstractNote={Abstract}, journal={BIOTECHNOLOGY JOURNAL}, author={Sternisha, Shawn M. and Mukherjee, Prabuddha and Alex, Aneesh and Chaney, Eric J. and Barkalifa, Ronit and Wan, Boyong and Lee, Jang Hyuk and Rico-Jimenez, Jose and Zurauskas, Mantas and Spillman, Darold R., Jr. and et al.}, year={2021}, month={Jun} }
 @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={Abstract}, 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} }