Enhanced cleaning strategies for UF/DF membranes in biopharmaceutical downstream processing

Abstract

Membrane fouling during ultrafiltration/diafiltration (UF/DF) remains a major challenge for extending membrane lifetime in biologics manufacturing. Developing cleaning strategies that combine high efficiency with scalability is essential for robust downstream processing. In this study, different UF/DF membrane cleaning methods were assessed under bench-scale conditions with consideration for large-scale implementation. Permeate-closed cleaning (PCC), which temporarily closes the permeate port to enhance shear, effectively mitigated normalized water permeability (NWP) decline. Furthermore, forward and reverse PCC at elevated feed flux achieved comparable fouling control with fewer steps by leveraging hydrodynamic shear and localized backwashing. To simplify large-scale adoption, high-flux PCC was developed and demonstrated the ability to restore NWP and maintain stability over extended cycles. Quantitatively, high‑flux PCC reduced early NWP decay by ~ 18% over the first three cycles and recovered > 15% NWP in previously fouled membranes, stabilizing permeability over extended reuse. Backwashing under controlled negative transmembrane pressure (TMP) further improved pore-level cleaning and extended membrane usability. Additionally, combining sodium hypochlorite (NaClO) with sodium hydroxide (NaOH) provided strong oxidative and alkaline action; within the evaluated range (25–150 ppm NaClO), 50 ppm was identified as a practical lower limit for effective recovery, while 150 ppm restored NWP from ~ 70 to ~ 100% with residual NaClO < 0.02 ppm after final rinsing. Collectively, these findings present a portfolio of cleaning strategies—hydrodynamic, chemical, and combined approaches—that enhance cleaning efficiency, reduce chemical exposure, and support sustainable UF/DF operations in biologics manufacturing.

Graphical abstract

Keywords

Ultrafiltration/diafiltration (UF/DF) / Membrane fouling / Cleaning strategies / Permeate-closed cleaning / Backwashing / Biologics downstream processing

References

[1] Ahmed A, Waghmare R, Kahn D, Wu Y (2006) Cleaning polyethersulfone membranes after ultrafiltration–diafiltration in monoclonal antibody production. BioPharm Int, 19(4), https://www.biopharminternational.com/view/cleaning-polyethersulfone-membranes-after-ultrafiltration-diafiltration-monoclonal-antibody-producti

[2] Arkhangelsky E, Kuzmenko D, Gitis V. Impact of chemical cleaning on properties and functioning of polyethersulfone membranes. J Membr Sci, 2007, 305(1–2): 176-184

[3] Baek Y, Singh N, Arunkumar A, Borys M, Li ZJ, Zydney AL. Ultrafiltration behavior of monoclonal antibodies and Fc-fusion proteins: effects of physical properties. Biotechnol Bioeng, 2017, 114(9): 2057-2065

[4] Baek Y, Yang D, Zydney AL. Development of a hydrodynamic cleaning cycle for ultrafiltration/diafiltration processes used for monoclonal antibody formulation. Ind Eng Chem Res, 2018, 57(47): 16110-16115

[5] Buss NA, Henderson SJ, McFarlane M, Shenton JM, de Haan L. Monoclonal antibody therapeutics: history and future. Curr Opin Pharmacol, 2012, 12(5): 615-622

[6] Carter PJ, Rajpal A. Designing antibodies as therapeutics. Cell, 2022, 185(15): 2789-2805

[7] Castro-Muñoz R, Cabezas R, Plata-Gryl M. Mangiferin: a comprehensive review on its extraction, purification and uses in food systems. Adv Colloid Interface Sci, 2024, 329 103188

[8] Chen J, Yu B, Cong H, Shen Y. Recent development and application of membrane chromatography. Anal Bioanal Chem, 2023, 415(1): 45-65

[9] Cheryan M (1998) Ultrafiltration and microfiltration handbook (2nd ed.). CRC Press. https://doi.org/10.1201/9781482278743

[10] Cui Z, Wang J, Zhang H, Ngo HH, Jia H, Guo W, Gao F, Yang G, Kang D. Investigation of backwashing effectiveness in membrane bioreactor (MBR) based on different membrane fouling stages. Bioresour Technol, 2018, 269: 355-362

[11] D’Souza NM, Mawson AJ. Membrane cleaning in the dairy industry: a review. Crit Rev Food Sci Nutr, 2005, 45(2): 125-134

[12] Desai M, Kundu A, Hageman M, Lou H, Boisvert D. Monoclonal antibody and protein therapeutic formulations for subcutaneous delivery: high-concentration, low-volume vs. low-concentration, high-volume. Mabs, 2023, 15(1): 2285277

[13] Ferreyra-Suarez D, Paredes-Vargas L, Jafari SM, García-Depraect O, Castro-Muñoz R. Extraction pathways and purification strategies towards carminic acid as natural-based food colorant: a comprehensive review. Adv Colloid Interface Sci, 2024, 323 103052

[14] Gitis V. Fouling and cleaning of ultrafiltration membranes in protein solution separation. J Water Process Eng, 2014,

[15] Hakimi MI, Farid MAA, Norrrahim MNF, Zakaria MR, Shirai Y, Hassan MA, Mohd Yusoff MZ. Tailored biochar from oil palm trunk via controlled carbonization for efficient dye adsorption. Bioresour Bioprocess, 2026, 13 24

[16] Huang S, Chen Y, Wang X, Guo J, Li Y, Dai L, Li S, Zhang S. Preparation of antifouling ultrafiltration membranes from copolymers of polysulfone and zwitterionic poly(arylene ether sulfone)s. Chin J Chem Eng, 2022, 49: 100-110

[17] Kim Y, Li S, Ghaffour N. Evaluation of different cleaning strategies for different types of forward osmosis membrane fouling and scaling. J Membr Sci, 2020, 596 117731

[18] Krack R. Chemical agents and costs in cleaning and disinfection of membrane equipment. Bull Int Dairy Fed, 1995, 4: 151-174

[19] Li L, Kumar S, Buck PM, Burns C, Lavoie J, Singh SK, Warne NW, Nichols P, Luksha N, Boardman D. Concentration dependent viscosity of monoclonal antibody solutions: explaining experimental behavior in terms of molecular properties. Pharm Res, 2014, 31(11): 3161-3178

[20] Li D, Gao C, Wang X, Wu G, Yin J, Huang Y, Sun X. Zwitterionic polysulfone copolymer/polysulfone blended ultrafiltration membranes with excellent thermostability and antifouling properties. Membranes, 2021, 11: 932

[21] Marino T, Blasi E, Tornaghi S, Di Nicolò E, Figoli A. Polyethersulfone membranes prepared with Rhodiasolv® Polarclean as water soluble green solvent. J Membr Sci, 2018, 549: 192-204

[22] Merck KGaA (2021) Pellicon® 3 Ultrafiltration Cassettes Installation and User Guide (MK_AN1065SEN00 Rev 7). 1–132. https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/310/065/pellicon3-installation-ug-an1065en-mk.pdf

[23] Merck KGaA (2026) Techniques for demonstrating cleaning effectiveness of ultrafiltration membranes (Technical Brief). Merck Millipore. https://www.merckmillipore.com/Web-ID-Site/id_ID/-/SGD/ShowDocument-Pronet?id=201512.119

[24] Mulyati S, Muchtar S, Arahman N, Syamsuddin Y, Mat Nawi NI, Harun NY, Bilad MR, Firdaus Y, Takagi R, Matsuyama H. Two-step dopamine-to-polydopamine modification of polyethersulfone ultrafiltration membrane for enhancing anti-fouling and ultraviolet resistant properties. Polymers, 2020, 12: 2051

[25] Niu H, Wang J, Liu M, Chai M, Zhao L, Liu X, Fan L, Tan W-S. Uridine modulates monoclonal antibody charge heterogeneity in Chinese hamster ovary cell fed-batch cultures. Bioresour Bioprocess, 2018, 5 42

[26] O’Mahony D, Bishop MR. Monoclonal antibody therapy. Front Biosci, 2006, 11: 1620-1635

[27] Ounifi I, Guesmi Y, Ursino C, Castro-Muñoz R, Agougui H, Jabli M, Hafiane A, Figoli A, Ferjani E. Synthesis and characterization of a thin-film composite nanofiltration membrane based on polyamide–cellulose acetate: application for water purification. J Polym Environ, 2022, 30: 707-718

[28] Pini Pereira P, Pacola Gonçalves I, Molina LCA, Delcolle R, Dzyazko YS, Moser Paraiso C, Batista Neto GL, Diório A, Marquetotti Salcedo Vieira A, Bergamasco R. Membrane for pressure‑driven separation prepared with a method of 3D printing: performance in concentrating orange peel extract. Membranes (Basel), 2025, 15(4): 105

[29] Rajendran SRC, Mason B, Doucette AA. Review of membrane separation models and technologies: processing complex food-based biomolecular fractions. Food Bioprocess Technol, 2021, 14: 415-428

[30] Samavedam R, Cote S, Morrison R (2007) Lifetime studies for membrane reuse: principles and case studies. BioPharm Int 20(9). https://www.biopharminternational.com/view/lifetime-studies-membrane-reuse-principles-and-case-studies

[31] Shepard HM, Phillips GL, D Thanos C, Feldmann M. Developments in therapy with monoclonal antibodies and related proteins. Clin Med, 2017, 17(3): 220-232

[32] Shi X, Tal G, Hankins NP, Gitis V. Fouling and cleaning of ultrafiltration membranes: a review. J Water Process Eng, 2014, 1: 121-138

[33] Vatanpour V, Yuksekdag A, Ağtaş M, Mehrabi M, Salehi E, Castro-Muñoz R, Koyuncu I. Zeolitic imidazolate framework (ZIF-8) modified cellulose acetate NF membranes for potential water treatment application. Carbohydr Polym, 2023, 299 120230

[34] WaterSep BioSeparations Corporation. (n.d.). Hollow Fiber Cleaning and Storage Procedure. https://www.researchgate.net/profile/Rafik-Karaman/post/How_do_I_clean_hollow_fiber/attachment/59d6262579197b80779846f0/AS%3A320442940755968%401453410958191/download/HF+SCleaning+Procedure.pdf

[35] Whitaker N, Pace SE, Merritt K, Tadros M, Khossravi M, Deshmukh S, Cheng Y, Joshi SB, Volkin DB, Dhar P. Developability assessments of monoclonal antibody candidates to minimize aggregation during large-scale ultrafiltration and diafiltration (UF-DF) processing. J Pharm Sci, 2022, 111(11): 2998-3008

[36] Wu Y, Ahmed A, et al. . Cleaning polyethersulfone membranes after ultrafiltration–diafiltration in monoclonal antibody production. BioPharm Int, 2006, 19(6): 67-74

[37] Yang L, Li D, Shi Y, Hill C, Pilliadugula R, et al. . High levels of microparticles release from biodegradable polylactic acid paper cups compared with polyethylene-lined cups. Chem Eng J, 2023, 468 143620

[38] Yang L, Qiu H, Lu Z, Zhang C, Liu G, Chang J, Wu C, Dong S, Yang S, Xiao F. Identification of performance and cost in a new backwash method to clean the UF membrane: backwashing with low dosage of NaClO. Environ Sci Pollut Res Int, 2023, 30(58): 121983-121992

[39] Yang J, Monnot M, Sun Y, Asia L, Wong-Wah-Chung P, Doumenq P, Moulin P. Microplastics in different water samples (seawater, freshwater, and wastewater): methodology approach for characterization using micro-FTIR spectroscopy. Water Res, 2023, 232 119711

[40] Zarghami S, Mohammadi T, Sadrzadeh M. Preparation, characterization and fouling analysis of in-air hydrophilic/underwater oleophobic bio-inspired polydopamine coated PES membranes for oily wastewater treatment. J Membr Sci, 2019,

RIGHTS & PERMISSIONS

The Author(s)