Moving from the bench towards a large scale, industrial platform process for adeno-associated viral vector purification.
adeno-associated virus (AAV)
downstream
gene therapy
purification
quality by design (QbD)
Journal
Biotechnology and bioengineering
ISSN: 1097-0290
Titre abrégé: Biotechnol Bioeng
Pays: United States
ID NLM: 7502021
Informations de publication
Date de publication:
10 2020
10 2020
Historique:
received:
07
05
2020
revised:
19
06
2020
accepted:
20
06
2020
pubmed:
24
6
2020
medline:
8
10
2021
entrez:
24
6
2020
Statut:
ppublish
Résumé
In recent years, there has been a strong interest in the development and production of gene therapy products, especially those utilizing adeno-associated virus (AAV) particles. This is evident with the growing number of clinical successes and agency approvals for AAV therapeutics. Due to this increased investment in this technology, a need exists for scalable commercial production methods to ensure adequate product supply as research in AAV shifts from bench-scale development to clinical production. The purpose of this review is to summarize current scalable purification techniques that can be employed during the commercial manufacturing of AAV as well as highlight certain development considerations, such as adventitious agent removal and process development using the principals of quality by design.
Types de publication
Journal Article
Review
Langues
eng
Sous-ensembles de citation
IM
Pagination
3199-3211Informations de copyright
© 2020 Wiley Periodicals LLC.
Références
Adamson-Small, L., Potter, M., Falk, D. J., Cleaver, B., Byrne, B. J., & Clément, N. (2016). A scalable method for the production of high-titer and high-quality adeno-associated type 9 vectors using the HSV platform. Molecular Therapy-Methods and Clinical Development, 3(March), 16031. https://doi.org/10.1038/mtm.2016.31
Alliance for Regenerative Medicine. (2019). 2019 Q3 quarterly regenerative medicine sector report. Retrieved from http://alliancerm.org/wp-content/uploads/2019/11/ARM_Q3_2019_FINAL-1.pdf
Alliance for Regenerative Medicine. (n.d.). Retrieved from https://alliancerm.org/manufacturing/
Anderson, R., Macdonald, I., Corbett, T., Whiteway, A., & Prentice, H. G. (2000). A method for the preparation of highly purified adeno-associated virus using affinity column chromatography, protease digestion and solvent extraction. Journal of Virological Methods, 85(1-2), 23-34. https://doi.org/10.1016/S0166-0934(99)00150-0
Atkinson, E. M., Takeya, R. K., & Aranha, I. L. (2003). Patent No. USOO6566118B1. Retrieved from https://patents.google.com/patent/US6566118B1/en
Auricchio, A., O'Connor, E., Hildinger, M., & Wilson, J. M. (2001). A single-step affinity column for purification of serotype-5 based adeno-associated viral vectors. Molecular Therapy, 4(4), 372-374. https://doi.org/10.1006/mthe.2001.0462
Barone, P. W., Wiebe, M. E., Leung, J. C., Hussein, I. T. M., Keumurian, F. J., Bouressa, J., … Springs, S. L. (2020). Viral contamination in biologic manufacture and implications for emerging therapies. Nature Biotechnology, 38, 563-572. https://doi.org/10.1038/s41587-020-0507-2
Bolton, G. R., & Mehta, K. K. (2016). The role of more than 40 years of improvement in protein A chromatography in the growth of the therapeutic antibody industry. Biotechnology Progress, 32(5), 1193-1202. https://doi.org/10.1002/btpr.2324
Burnham, B., Nass, S., Kong, E., Mattingly, M. E., Woodcock, D., Song, A., … O'Riordan, C. R. (2015). Analytical ultracentrifugation as an approach to characterize recombinant adeno-associated viral vectors. Human Gene Therapy Methods, 26(6), 228-242. https://doi.org/10.1089/hgtb.2015.048
Burova, E., & Ioffe, E. (2005). Chromatographic purification of recombinant adenoviral and adeno-associated viral vectors: Methods and implications. Gene Therapy, 12, S5-S17. https://doi.org/10.1038/sj.gt.3302611
Chahal, P. S., Aucoin, M. G., & Kamen, A. (2007). Primary recovery and chromatographic purification of adeno-associated virus type 2 produced by baculovirus/insect cell system. Journal of Virological Methods, 139(1), 61-70. https://doi.org/10.1016/j.jviromet.2006.09.011
Clément, N., & Grieger, J. C. (2016). Manufacturing of recombinant adeno-associated viral vectors for clinical trials. Molecular Therapy-Methods and Clinical Development, 3(September), 16002. https://doi.org/10.1038/mtm.2016.2
Crosson, S. M., Dib, P., Smith, J. K., & Zolotukhin, S. (2018). Helper-free production of laboratory grade AAV and purification by iodixanol density gradient centrifugation. Molecular Therapy-Methods and Clinical Development, 10(September), 1-7. https://doi.org/10.1016/j.omtm.2018.05.001
Davidoff, A. M., Ng, C. Y. C., Sleep, S., Gray, J., Azam, S., Zhao, Y., … Nathwani, A. C. (2004). Purification of recombinant adeno-associated virus type 8 vectors by ion exchange chromatography generates clinical grade vector stock. Journal of Virological Methods, 121(2), 209-215. https://doi.org/10.1016/j.jviromet.2004.07.001
Dias Florencio, G., Precigout, G., Beley, C., Buclez, P. O., Garcia, L., & Benchaouir, R. (2015). Simple downstream process based on detergent treatment improves yield and in vivo transduction efficacy of adeno-associated virus vectors. Molecular Therapy-Methods and Clinical Development, 2(May), 15024. https://doi.org/10.1038/mtm.2015.24
Dismuke, D., Tenenbaum, L., & Samulski, R. (2013). Biosafety of recombinant adeno-associated virus vectors. Current Gene Therapy, 13(6), 434-452. https://doi.org/10.2174/15665232113136660007
Dismuke, D. J., & Kotin, R. M. (2017). Obstacles for rAAV clinical trials: A question of vector supply and demand or know-how. Cell and Gene Therapy Insights, 3(9), 755-768. https://doi.org/10.18609/cgti.2017.075
FDA. (2018). Chemistry, manufacturing, and control (CMC) for human gene therapy investigational new drug applications (INDs). Retrieved from http://www.fda.org
FDA. (2019). Statement from FDA Commissioner Scott Gottlieb, M.D. and Peter Marks, M.D., Ph.D., Director of the Center for Biologics: Evaluation and Research on new policies to advance development of safe and effective cell and gene therapies. Retrieved from FDA Statement website: https://www.fda.gov/news-events/press-announcements/statement-fda-commissioner-scott-gottlieb-md-and-peter-marks-md-phd-director-center-biologics
Forsberg, N., Glover, C., Hughes, J., Jaluria, P., King, D., Madsen, J., & Alan, M. (2018). Key considerations in gene therapy manufacturing for commercialization the future of medicine: Gene therapy makes a compelling case. Cell Culture Dish. Retrieved from https://cellculturedish.com/wp-content/uploads/2018/09/Key-Considerations-Gen-Therapy-Manufacturing-Commercialization.pdf
Gagnon, P., Makovsek, V., Stokelj, M., Jug, H., Zigon, R., Batic, L., … Strancar, A. (2019). Cornerstone technology for purification of clinical-quality AAV. BPI Cell and Gene Therapy Conference.
GE Healthcare Life Sciences. (2016). AVB sepharose high performance product manual. Product Manual. Retrieved from https://www.gelifesciences.com/gehcls_images/GELS/RelatedContent/Files/1481286259681/litdoc28920754_20161209132411.pdf
GE Healthcare Life Sciences. (2018a). CaptoTM AVB Product Manual. Product Manual. Retrieved from https://cdn.gelifesciences.com/dmm3bwsv3/AssetStream.aspx?mediaformatid=10061&destinationid=10016&assetid=27081
GE Healthcare Life Sciences. (2018b). Optimization of midstream cell lysis and virus filtration steps in an adenovirus purification process. Retrieved from https://cdn.gelifesciences.com/dmm3bwsv3/AssetStream.aspx?mediaformatid=10061&destinationid=10016&assetid=26781
Grimm, D., Kern, A., Rittner, K., & Kleinschmidt, J. (1998). Novel tool for production and purification recombinant adenoassociated virus vectors. Human Gene Therapy, 9, 2745-2760.
Gruntman, A. M., Su, L., Su, Q., Gao, G., Mueller, C., & Flotte, T. R. (2015). Stability and compatibility of recombinant adeno-associated virus under conditions commonly encountered in human gene therapy trials. Human Gene Therapy Methods, 26(2), 71-76. https://doi.org/10.1089/hgtb.2015.040
Gu, B., Bhat, V., Dong, W., Pham, H., Pubill, S., Kasaraneni, N., … Seth, A. (2018). Establishment of a scalable manufacturing platform for in-silico- derived ancestral adeno-associated virus vectors. Cell and Gene Therapy Insights, 4(S1), 753-769. https://doi.org/10.18609/cgti.2018.079
Guo, P., El-Gohary, Y., Prasadan, K., Shiota, C., Xiao, X., Wiersch, J., … Gittes, G. K. (2012). Rapid and simplified purification of recombinant adeno-associated virus. Journal of Virological Methods, 183(2), 139-146. https://doi.org/10.1016/j.jviromet.2012.04.004
Hebben, M. (2018). Downstream bioprocessing of AAV vectors: Industrial challenges & regulatory requirements. Cell and Gene Therapy Insights, 4(2), 131-146. https://doi.org/10.18609/cgti.2018.016
Heldt, C. L. (2019). Scalable method utilizing low pH for DNA removal in the harvest of recombinant adeno-associated virus vectors. Journal of Chromatography B, 1124(June), 173-179. https://doi.org/10.1016/j.jchromb.2019.06.011
Hermens, W. T. J. M. C., & Smith, J. P. (2013). Patent No. WO 2013036118.
ICH. (1999). Viral safety evaluation of biotechnology products derived from cell lines of human or animal origin: Q5A(R1). Retrieved from http://www.ich.org/
ICH. (2009). ICH harmonised tripartite guideline: Pharmaceutical development: Q8 (R2). Retrieved from http://www.ich.org/
ICH. (2019). Technical and regulatory considerations for pharmaceutical product lifecycle management: Q12. Retrieved from http://www.ich.org/
Khanal, O., Singh, N., Traylor, S. J., Xu, X., Ghose, S., Li, Z. J., & Lenhoff, A. M. (2018). Contributions of depth filter components to protein adsorption in bioprocessing. Biotechnology and Bioengineering, 115(8), 1938-1948. https://doi.org/10.1002/bit.26707
Kimura, T., Ferran, B., Tsukahara, Y., Shang, Q., Desai, S., Fedoce, A., … Bachschmid, M. M. (2019). Production of adeno-associated virus vectors for in vitro and in vivo applications. Scientific Reports, 9(1), 1-13. https://doi.org/10.1038/s41598-019-49624-w
Lerch, T. F., & Chapman, M. S. (2012). Identification of the heparin binding site on adeno-associated virus serotype 3B (AAV-3B). Virology, 423(1), 6-13. https://doi.org/10.1016/j.virol.2011.10.007
Leskovec, M., Primec, S., Gagnon, P., & Štrancar, A. (2018). An Industrial Purification Platform For AAV. Retrieved from https://www.biaseparations.com/en/library/posters/988/an-industrial-purification-platform-for-aav
Lock, M., Alvira, M. R., & Wilson, J. M. (2012). Analysis of particle content of recombinant adeno-associated virus serotype 8 vectors by ion-exchange chromatography. Human Gene Therapy Methods, 23(1), 56-64. https://doi.org/10.1089/hgtb.2011.217
Lundgren, M. (2020). A scalable AAV production process from cell culture to purified bulk. Presented at ASGCT 23rd annual meeting, Virtual Meeting.
Maheshwari, G., Jannat, R., McCormick, L., & Hsu, D. (2004). Thermal inactivation of adenovirus type 5. Journal of Virological Methods, 118(2), 141-146. https://doi.org/10.1016/j.jviromet.2004.02.003
Mattila, J., Clark, M., Liu, S., Pieracci, J., Gervais, T. R., Wilson, E., … Simpson-Platre, C. (2016). Retrospective evaluation of Low-pH viral inactivation and viral filtration data from a multiple company collaboration. PDA Journal of Pharmaceutical Science and Technology, 70(3), 293-299. https://doi.org/10.5731/pdajpst.2016.006478
Mattila, J., Curtis, S., Webb-Vargas, Y., Wilson, E., Galperina, O., Roush, D., … Bennett, L. (2019). Retrospective evaluation of cycled resin in viral clearance studies-A multiple company collaboration. PDA Journal of Pharmaceutical Science and Technology, 73(5), 470-486. https://doi.org/10.5731/pdajpst.2018.009605
Miesegaes, G. R., Lute, S. C., Read, E. K., & Brorson, K. A. (2014). Viral clearance by flow-through mode ion exchange columns and membrane adsorbers. Biotechnology Progress, 30(1), 124-131. https://doi.org/10.1002/btpr.1832
Millipore Sigma. (2016). Benzonase® Endonuclease Brochure. Retrieved from https://www.emdmillipore.com/US/en
Naso, M. F., Tomkowicz, B., Perry, W. L., & Strohl, W. R. (2017). Adeno-associated virus (AAV) as a vector for gene therapy. BioDrugs, 31(4), 317-334. https://doi.org/10.1007/s40259-017-0234-5
Nass, S. A., Mattingly, M. A., Woodcock, D. A., Burnham, B. L., Ardinger, J. A., Osmond, S. E., … O'Riordan, C. R. (2018). Universal Method for the Purification of Recombinant AAV Vectors of Differing Serotypes. Molecular Therapy - Methods and Clinical Development, 9(June), 33-46. https://doi.org/10.1016/j.omtm.2017.12.004
Nguyen, H. C., Langland, A. L., Amara, J. P., Dullen, M., Kahn, D. S., & Costanzo, J. A. (2019). Improved HCP Reduction Using a New, All-Synthetic Depth Filtration Media Within an Antibody Purification Process. Biotechnology Journal, 14(1), 1-11. https://doi.org/10.1002/biot.201700771
Nian, R., Zhang, W., Tan, L., Lee, J., Bi, X., Yang, Y., … Gagnon, P. (2016). Advance chromatin extraction improves capture performance of protein A affinity chromatography. Journal of Chromatography A, 1431, 1-7. https://doi.org/10.1016/j.chroma.2015.12.044
Okada, T., Nonaka-Sarukawa, M., Uchibori, R., Kinoshita, K., Hayashita-Kinoh, H., Nitahara-Kasahara, Y., … Ozawa, K. (2009). Scalable Purification of Adeno-Associated Virus Serotype 1 (AAV1) and AAV8 Vectors, Using Dual Ion-Exchange Adsorptive Membranes. Human Gene Therapy, 20, 1013-1021. https://doi.org/10.1089/hum.2009.006
Paulene, M.Q., S., Gagnon, P., Nichols, G., & Thorne, B. A. (2010). Patent No. WO 2010148143.
Penaud-Budloo, M., François, A., Clément, N., & Ayuso, E. (2018). Pharmacology of Recombinant Adeno-associated Virus Production. Molecular Therapy: Methods & Clinical Development, 8(March), 166-180. https://doi.org/10.1016/j.omtm.2018.01.002
Pierson, E. E., Keifer, D. Z., Asokan, A., & Jarrold, M. F. (2016). Resolving Adeno-Associated Viral Particle Diversity with Charge Detection Mass Spectrometry. Analytical Chemistry, 88(13), 6718-6725. https://doi.org/10.1021/acs.analchem.6b00883
Piras, B. A., Drury, J. E., Morton, C. L., Spence, Y., Lockey, T. D., Nathwani, A. C., … Meagher, M. M. (2016). Distribution of AAV8 particles in cell lysates and culture media changes with time and is dependent on the recombinant vector. Molecular Therapy-Methods and Clinical Development, 3(February), 16015. https://doi.org/10.1038/mtm.2016.15
Potter, M., Lins, B., Mietzsch, M., Heilbronn, R., Van Vliet, K., Chipman, P., … Zolotukhin, S. (2014). A simplified purification protocol for recombinant adeno-associated virus vectors. Molecular Therapy - Methods and Clinical Development, 1(March), 14034. https://doi.org/10.1038/mtm.2014.34
Prazeres, D. M. F. (2016). Considerations on the Use of Enzymes in the Downstream Processing of Biopharmaceuticals. Pharmaceutical Bioprocessing, 4(5), 95-99.
Qu, G., Bahr-Davidson, J., Prado, J., Tai, A., Cataniag, F., Mcdonnell, J., … Wright, J. F. (2007). Separation of adeno-associated virus type 2 empty particles from genome containing vectors by anion-exchange column chromatography. Journal of Virological Methods, 140, 183-192. https://doi.org/10.1016/j.jviromet.2006.11.019
Qu, W., Wang, M., Wu, Y., & Xu, R. (2015). Scalable downstream strategies for purification of recombinant adeno- associated virus vectors in light of the properties. Current Pharmaceutical Biotechnology, 16(8), 684-695. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/25941887%0A, http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid
Raghavan, B., Collins, M., Walls, S., Lambropoulos, A., & Bergheim-Pietza, S. (2019). Optimizing the clarification of industrial scale viral vector culture for gene therapy. Cell and Gene Therapy Insights, 5(9), 1311-1322. https://doi.org/10.18609/cgti.2019.137
Rathore, A. S. (2009). Roadmap for implementation of quality by design (QbD) for biotechnology products. Trends in Biotechnology, 27(9), 546-553. https://doi.org/10.1016/j.tibtech.2009.06.006
Rayaprolu, V., Kruse, S., Kant, R., Venkatakrishnan, B., Movahed, N., Brooke, D., … Bothner, B. (2013). Comparative analysis of adeno-associated virus capsid stability and dynamics. Journal of Virology, 87(24), 13150-13160. https://doi.org/10.1128/jvi.01415-13
Reth, M. (2013). Matching cellular dimensions with molecular sizes. Nature Immunology, 14(8), 765-767. https://doi.org/10.1038/ni.2621
Ruppach, H. (2014). Log10 reduction factors in viral clearance studies. BioProcessing Journal, 12(4), 24-30. https://doi.org/10.12665/j124.ruppach
Sharnez, R., Doares, S., Manning, S., Mehta, K., Mahajan, E., To, A., … Tjandra, H. (2018). Multiproduct resin reuse for clinical and commercial manufacturing-Methodology and acceptance criteria. PDA Journal of Pharmaceutical Science and Technology, 72(6), 584-598. https://doi.org/10.5731/pdajpst.2016.007245
Shuhei Sakamoto, K., Yashuhiro Kawano, M., Katsuyuki Dodo, K., Tatsuji Enoki, K., Hirofumi Yoshioka, R., Hikaru Takakura, K., & Junichi Mineno, U. (2018). Patent No. US10072250B2.
Smith, J., Grieger, J. C., & Samulski, R. J. (2018). Overcoming bottlenecks in AAV manufacturing for gene therapy. Cell and Gene Therapy Insights, 4(8), 815-825. https://doi.org/10.18609/cgti.2018.083
Smith, P. H., Wright, J. F., Qu, G., Parker, A., & Sommer, J. M. (2003). 901. Evidence that ionic interactions are involved in concentration-induced aggregation of recombinant adeno-associated virus. Molecular Therapy, 7(5), S348. https://doi.org/10.1016/S1525-0016(16)41343-2
Smith, R. H., Levy, J. R., & Kotin, R. M. (2009). A simplified baculovirus-AAV expression vector system coupled with one-step affinity purification yields high-titer rAAV stocks from insect cells. Molecular Therapy, 17(11), 1888-1896. https://doi.org/10.1038/mt.2009.128
Sommer, J. M., Smith, P. H., Parthasarathy, S., Isaacs, J., Vijay, S., Kieran, J., … Wright, F. (2003). Quantification of adeno-associated virus particles and empty capsids by optical density measurement. Molecular Therapy, 7(1), 122-128. https://doi.org/10.1016/S1525-0016(02)00019-9
Sung, Y. K., & Kim, S. W. (2019). Recent advances in the development of gene delivery systems. Biomaterials Research, 23(1), 1-7. https://doi.org/10.1186/s40824-019-0156-z
Terova, O., Soltys, S., Hermans, P., De Rooij, J., & Detmers, F. (2018). Overcoming downstream purification challenges for viral vector manufacturing: Enabling advancement of gene therapies in the clinic. Cell and Gene Therapy Insights, 4(2), 101-111. https://doi.org/10.18609/cgti.2018.017
Thermo Fisher Scientific. (2012). POROSTM CaptureSelectTM AAV Resins: AAV8, AAV9, AAVX. User Guide. Retrieved from https://assets.thermofisher.com/TFS-Assets/LSG/manuals/100038399_POROS_CapSel_AAV8_AAV9_Resins_UG.pdf
Thorne, B. A., Quigley, P., Nichols, G., Moore, C., Pastor, E., Price, D., … Peluso, R. W. (2008). Characterizing clearance of helper adenovirus by a clinical rAAV1 manufacturing process. Biologicals, 36(1), 7-18. https://doi.org/10.1016/j.biologicals.2007.04.001
Tomono, T., Hirai, Y., Okada, H., Miyagawa, Y., Adachi, K., Sakamoto, S., … Okada, T. (2018). Highly efficient ultracentrifugation-free chromatographic purification of recombinant AAV serotype 9. Molecular Therapy-Methods and Clinical Development, 11(December), 180-190. https://doi.org/10.1016/j.omtm.2018.10.015
Toueille, M. (2018). Development of purification steps for several AAV serotypes using POROSTM CaptureSelectTM AAVX affinity chromatography. Cell and Gene Therapy Insights, 4(7), 637-645. https://doi.org/10.18609/cgti.2018.061
Urabe, M., Xin, K. Q., Obara, Y., Nakakura, T., Mizukami, H., Kume, A., … Ozawa, K. (2006). Removal of empty capsids from type 1 adeno-associated virus vector stocks by anion-exchange chromatography potentiates transgene expression. Molecular Therapy, 13(4), 823-828. https://doi.org/10.1016/j.ymthe.2005.11.024
Vandenberghe, L. H., Xiao, R., Lock, M., Lin, J., Korn, M., & Wilson, J. M. (2010). Efficient serotype-dependent release of functional vector into the culture medium during adeno-associated virus manufacturing. Human Gene Therapy, 21(10), 1251-1257. https://doi.org/10.1089/hum.2010.107
Wang, C., Hari, S., Mulagapati, R., Chen, Z., Du, J., Zhao, X., … Liu, D. (2019). Developing an anion exchange chromatography assay for determining empty and full capsid contents in AAV6.2. Molecular Therapy: Methods & Clinical Development, 15(December), 257-263. https://doi.org/10.1016/j.omtm.2019.09.006
Wang, D., Tai, P. W. L., & Gao, G. (2019). Adeno-associated virus vector as a platform for gene therapy delivery. Nature Reviews Drug Discovery, 18(5), 358-378. https://doi.org/10.1038/s41573-019-0012-9
Wang, Q., Lock, M., Prongay, A. J., Alvira, M. R., Petkov, B., & Wilson, J. M. (2015). Identification of an adeno-associated virus binding epitope for AVB sepharose affinity resin. Molecular Therapy-Methods and Clinical Development, 2(July), 15040. https://doi.org/10.1038/mtm.2015.40
Ward, A. (2018). Exploring a new enzymatic tool for AAV production. Genetic Engineering and Biotechnology News, 38(3), 22. https://doi.org/10.1089/gen.38.03.10
Wright, J. F. (2008). Manufacturing and characterizing AAV-based vectors for use in clinical studies. Gene Therapy, 15(11), 840-848. https://doi.org/10.1038/gt.2008.65
Wright, J. F., Le, T., Prado, J., Bahr-Davidson, J., Smith, P. H., Zhen, Z., … Qu, G. (2005). Identification of factors that contribute to recombinant AAV2 particle aggregation and methods to prevent its occurrence during vector purification and formulation. Molecular Therapy, 12(1), 171-178. https://doi.org/10.1016/j.ymthe.2005.02.021
Wright, J. F., & Qu, G. (2006). Patent No. US20170247664A1.
Wu, Z., Asokan, A., & Samulski, R. J. (2006). Adeno-associated virus serotypes: Vector toolkit for human gene therapy. Molecular Therapy, 14(3), 316-327. https://doi.org/10.1016/j.ymthe.2006.05.009
Ye, G. jie, Scotti, M. M., Thomas, D. L., Wang, L., Knop, D. R., & Chulay, J. D. (2014). Herpes simplex virus clearance during purification of a recombinant adeno-associated virus serotype 1 vector. Human Gene Therapy Clinical Development, 25(4), 212-217. https://doi.org/10.1089/humc.2014.060
Zhang, G.-H., Baek, L., Buchardt, O., & Koch, C. (1994). Differential blocking of coagulation-activating pathways of Limulus amebocyte lysate. Journal of Clinical Microbiology, 32(6), 1537-1541. https://doi.org/10.1128/jcm.32.6.1537-1541.1994
Zhang, L., & Mao, S. (2017). Application of quality by design in the current drug development. Asian Journal of Pharmaceutical Sciences, 12(1), 1-8. https://doi.org/10.1016/j.ajps.2016.07.006
Zolotukhin, S., Byrne, B. J., Mason, E., Zolotukhin, I., Potter, M., Chesnut, K., … Muzyczka, N. (1999). Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Therapy, 6(6), 973-985. https://doi.org/10.1038/sj.gt.3300938