@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} }
 @misc{bacon_lavoie_rao_daniele_menegatti_2020, title={Past, Present, and Future of Affinity-based Cell Separation Technologies}, volume={112}, ISSN={["1878-7568"]}, DOI={10.1016/j.actbio.2020.05.004}, abstractNote={Progress in cell purification technology is critical to increase the availability of viable cells for therapeutic, diagnostic, and research applications. A variety of techniques are now available for cell separation, ranging from non-affinity methods such as density gradient centrifugation, dielectrophoresis, and filtration, to affinity methods such as chromatography, two-phase partitioning, and magnetic-/fluorescence-assisted cell sorting. For clinical and analytical procedures that require highly purified cells, the choice of cell purification method is crucial, since every method offers a different balance between yield, purity, and bioactivity of the cell product. For most applications, the requisite purity is only achievable through affinity methods, owing to the high target specificity that they grant. In this review, we discuss past and current methods for developing cell-targeting affinity ligands and their application in cell purification, along with the benefits and challenges associated with different purification formats. We further present new technologies, like stimuli-responsive ligands and parallelized microfluidic devices, towards improving the viability and throughput of cell products for tissue engineering and regenerative medicine. Our comparative analysis provides guidance in the multifarious landscape of cell separation techniques and highlights new technologies that are poised to play a key role in the future of cell purification in clinical settings and the biotech industry. STATEMENT OF SIGNIFICANCE: Technologies for cell purification have served science, medicine, and industrial biotechnology and biomanufacturing for decades. This review presents a comprehensive survey of this field by highlighting the scope and relevance of all known methods for cell isolation, old and new alike. The first section covers the main classes of target cells and compares traditional non-affinity and affinity-based purification techniques, focusing on established ligands and chromatographic formats. The second section presents an excursus of affinity-based pseudo-chromatographic and non-chromatographic technologies, especially focusing on magnetic-activated cell sorting (MACS) and fluorescence-activated cell sorting (FACS). Finally, the third section presents an overview of new technologies and emerging trends, highlighting how the progress in chemical, material, and microfluidic sciences has opened new exciting avenues towards high-throughput and high-purity cell isolation processes. This review is designed to guide scientists and engineers in their choice of suitable cell purification techniques for research or bioprocessing needs.}, journal={ACTA BIOMATERIALIA}, author={Bacon, Kaitlyn and Lavoie, Ashton and Rao, Balaji M. and Daniele, Michael and Menegatti, Stefano}, year={2020}, month={Aug}, pages={29–51} }
 @article{saberi-bosari_omary_lavoie_prodromou_day_menegatti_san-miguel_2019, title={Affordable Microfluidic Bead-Sorting Platform for Automated Selection of Porous Particles Functionalized with Bioactive Compounds}, volume={9}, ISSN={["2045-2322"]}, url={http://dx.doi.org/10.1038/s41598-019-42869-5}, DOI={10.1038/s41598-019-42869-5}, abstractNote={Abstract}, number={1}, journal={SCIENTIFIC REPORTS}, publisher={Springer Science and Business Media LLC}, author={Saberi-Bosari, Sahand and Omary, Mohammad and Lavoie, Ashton and Prodromou, Raphael and Day, Kevin and Menegatti, Stefano and San-Miguel, Adriana}, year={2019}, month={May} }
 @article{day_prodromou_bosari_lavoie_omary_market_san miguel_menegatti_2019, title={Discovery and Evaluation of Peptide Ligands for Selective Adsorption and Release of Cas9 Nuclease on Solid Substrates}, volume={30}, ISSN={["1520-4812"]}, url={http://dx.doi.org/10.1021/acs.bioconjchem.9b00703}, DOI={10.1021/acs.bioconjchem.9b00703}, abstractNote={The rapid expansion of CRISPR in biotechnology, medicine, and bioprocessing poses an urgent need for advanced manufacturing of Cas nucleases. The lack of Cas-targeting ligands, however, prevents the development of platform processes for purifying this class of molecules. This work represents the first effort at developing short synthetic Cas9-binding peptides and demonstrates their applicability as affinity ligands for the purification of a Cas nuclease. Candidate Cas9-targeting peptides were initially identified by screening a solid-phase peptide library against a model mixture of Streptococcus pyogenes Cas9 spiked in Escherichia coli cell lysate. An ensemble of homologous sequences were identified, conjugated on Toyopearl resin, and evaluated by Cas9 binding studies to identify sequences providing selective Cas9 capture and efficient release. In silico docking studies were also performed to evaluate the binding energy and site of the various peptides on Cas9. Notably, sequences GYYRYSEY and YYHRHGLQ were shown to target the RecII domain of Cas9, which is not involved in nuclease activity, and was targeted as ideal binding site. The peptide ligands were validated by purifying Cas9 from the E. coli lysate in dynamic conditions and through measurements of binding capacity and strength (Qmax and KD). The resulting values of Qmax = 4 - 5 mg Cas9 per mL of resin and KD ~ 0.1 - 0.3 μM, and product recovery (86 - 89%) and purity (91% - 93%) indicate that both peptides, and YYHRHGLQ in particular, can serve as capture ligands in a platform purification process of Cas9.}, number={12}, journal={BIOCONJUGATE CHEMISTRY}, publisher={American Chemical Society (ACS)}, author={Day, Kevin and Prodromou, Raphael and Bosari, Sahand Saberi and Lavoie, Ashton and Omary, Mohammad and Market, Connor and San Miguel, Adriana and Menegatti, Stefano}, year={2019}, month={Dec}, pages={3057–3068} }
 @article{lavoie_fazio_blackburn_goshe_carbonell_menegatti_2019, title={Targeted Capture of Chinese Hamster Ovary Host Cell Proteins: Peptide Ligand Discovery}, volume={20}, ISSN={["1422-0067"]}, DOI={10.3390/ijms20071729}, abstractNote={The growing integration of quality-by-design (QbD) concepts in biomanufacturing calls for a detailed and quantitative knowledge of the profile of impurities and their impact on the product safety and efficacy. Particularly valuable is the determination of the residual level of host cell proteins (HCPs) secreted, together with the product of interest, by the recombinant cells utilized for production. Though often referred to as a single impurity, HCPs comprise a variety of species with diverse abundance, size, function, and composition. The clearance of these impurities is a complex issue due to their cell line to cell line, product-to-product, and batch-to-batch variations. Improvements in HCP monitoring through proteomic-based methods have led to identification of a subset of “problematic” HCPs that are particularly challenging to remove, both at the product capture and product polishing steps, and compromise product stability and safety even at trace concentrations. This paper describes the development of synthetic peptide ligands capable of capturing a broad spectrum of Chinese hamster ovary (CHO) HCPs with a combination of peptide species that allow for advanced mixed-mode binding. Solid phase peptide libraries were screened for identification and characterization of peptides that capture CHO HCPs while showing minimal binding of human IgG, utilized here as a model product. Tetrameric and hexameric ligands featuring either multipolar or hydrophobic/positive amino acid compositions were found to be the most effective. Tetrameric multipolar ligands exhibited the highest targeted binding ratio (ratio of HCP clearance over IgG loss), more than double that of commercial mixed-mode and anion exchange resins utilized by industry for IgG polishing. All peptide resins tested showed preferential binding to HCPs compared to IgG, indicating potential uses in flow-through mode or weak-partitioning-mode chromatography.}, number={7}, journal={INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES}, author={Lavoie, R. Ashton and Fazio, Alice and Blackburn, R. Kevin and Goshe, Michael B. and Carbonell, Ruben G. and Menegatti, Stefano}, year={2019}, month={Apr} }