Cleanascite™ Lipid Removal Reagent and Clarification

Cell Response

Suarez-Carmona,Meggy, et al. "Harnessinglipid-driven immunometabolic pathways in omental metastases toenhance immunotherapy in patients with ovarian cancer."Signal Transduction andTargeted Therapy 11.1(2026): 78. The article states “Similar to maraviroc and anti-CD36treatment, depletinglipids from ascitesassociated macrophages (AAMs) by ‘skimming’ the ascitessupernatants with Cleanascite™led to a striking increase in cytokine production, indicative ofmacrophage activation.”

Plasma/Serum

Post-StrokeImmune Suppression is Mediated by Bioactive Lipids ”,An abstract describes the use of Cleanascite™ to compare stroke plasma, in ex vivo models of immune suppression.

Tumormicroenvironment (TME)

Dyer A.,Dudley R., Ahuja S. et al. Characterisationof human in vitro tumour-associated macrophage models to definetranslational relevance . Sci Rep (2025). https://doi.org/10.1038/ s41598-025-30224-w. Depletion of lipidswith Cleanascite™ from tumor conditioned media confirmed that lipidfactors are partly responsiblefor the polarization of Tumor Associated Macrophages (TAMs) towards asuppressive phenotype.

Dalangood,Sumiya et al. Cancer-associatedadipocytes mediate CD8 + T cell dysfunction via FGF21-drivenlipolysis CellReports, Volume 44, Issue 11, 116526. Through the selective depletionof lipids with Cleanascite™,it was determined that fatty acids released by cancer-associatedadipocytes (CAAs) are critical for CD8 + T cell exhaustion in thetumor microenvironment.

Curtin,Meghan C., et al. "Leanbreast adipocytes secrete an oxylipin that suppresses breast cancervia ferroptosis ." bioRxiv (2025):2025-07. Using Cleanascite™ for selective lipid depletion, the study pinpointed a bioactiveoxylipin as a keytumor-suppressive factor.

Chu,Tianhao, et al. "Metabolismarchetype cancer cells induce protumor TREM2+ macrophages viaoxLDL-mediated metabolic interplay in hepatocellularcarcinoma. " NatureCommunications 16.1(2025): 6770. Cleanascite™ lipid depletion abolished the induction of TREM2 and SPP1 inmacrophages by tumor interstitial fluid (TIF), confirming that oxLDLmediates TAM polarization through the TREM2–SYK–CEBPα pathway.

Li,Xiaoyun, et al. " Tumor-instructedglutamine synthesis in cancer-associated fibroblasts promotespro-tumor macrophages." Journalof Experimental Medicine 222.9(2025): e20241426. Lipid-depleted Tumor-conditioned media derivedfrom Cleanascite™ significantly downregulated IL-6 and glutamine synthesis pathwaygenes in fibroblasts, impairing their ability to promote TAMpolarization and immunosuppression.

Slattery,Karen, et al. " Uptakeof lipids from ascites drives NK cell metabolic dysfunction inovarian cancer. " ScienceImmunology 10.107(2025): eadr4795. The article states: “To test whether uptake ofpolar lipids drives NK cell dysfunction in ascites, we depleted ascites of lipids using a lipid adsorption and clarificationreagent (Cleanascite™) which caused widespread lipid depletion inascites”. Removinglipids from ascites abrogated the suppression of the immune system byascites.

Gurunget al., “Stromal lipid species dictate melanoma metastasis andtropism” 2025, Cancer Cell 43, 1108–1124 June 9, 2025 https://doi.org/10.1016/j.ccell.2025.04.001. Thearticle states “To verify lipids in YA secretomes lead to increasedOXPHOS, wedelipidated YA and OA secretomes with Cleanascite, beforetransferring the secretome to melanoma cells. This work adds to the growing evidence that extrinsic lipid cues fromthe microenvironment shape tumor cell behavior.

CellResponse

Toraason, Erik, et al. Caenorhabditiselegans avoids Todstoff, a novel nociceptive necrotaxiscue " bioRxiv (2026):2026-02. In part through the use of Cleanascite™,a novel mode of post-mortem inter-animal chemical communication wasuncovered to promote death avoidance.


Flury,Anna, et al. " Aneurodegenerative cellular stress response linked to dark microgliaand toxic lipid secretio n." Neuron (2024).

Thebrain’s primary immune cells, microglia, are a leading causal celltype in Alzheimer’s disease (AD). Yet, the mechanisms by whichmicroglia can drive neurodegeneration remain unresolved. Theintegrated stress response (ISR), characterizes a microglia subsetwith neurodegenerative outcomes. Mechanistically, evidence ispresented that ISR activation promotes the secretion of toxic lipidsby microglia, impairing neuron homeostasis and survival invitro .Notably, eliminating lipids using an adsorbent resin (Cleanascite)abolished these toxic effects.

Dahal,Ankit, et al. " Platelet-activatingfactor (PAF) promotes immunosuppressive neutrophil differentiationwithin tumors ." Proceedingsof the National Academy of Sciences 121.35(2024): e2406748121.

Chronicinflammatory milieu in the tumor microenvironment (TME) leads to therecruitment and differentiation of myeloid- derived suppressor cells(MDSCs). Polymorphonuclear (PMN)- MDSCs, which are phenotypically andmorphologically defined as a subset of neutrophils, cause majorimmune suppression in the TME. Despite recent advances in ourunderstanding of PMN- MDSC functions, the mechanism that gives riseto immunosuppressive neutrophils within the TME remains elusive. Totest the hypothesis that cancer-secreted mediators were lipids, weselectively depleted lipids from the cancer cell supernatant using alipid-binding micelle kit (Cleanascite™) .

Wang,H., Yung, M.M., Xuan, Y. etal. Polyunsaturatedfatty acids promote M2-like TAM deposition via dampening RhoA-YAP1signaling in the ovarian cancer microenvironment . ExpHematol Oncol 13,90 (2024). https://doi.org/10.1186/s40164-024-00558-8.Tumor-associated-macrophages (TAMs) are multi-polarized. However, theimpact of omental conditioned medium/ascites (OCM/AS) on TAMpolarization and its function in tumor progression remains elusive.The article states “Recent studies have suggested that lipidaccumulation and metabolism are associated with the differentiationand activation of protumoral TAMs. Hence, we examined the lipiddroplets (LDs) in MФs, which are cellular storage organelles forneutral lipids. … theremoval of free fatty acids in OCM by Cleanascite™ attenuatedLDs deposition in OCM-C MФs ,indicating M2 MФs and OCM-MФs exhibited higher lipid accumulationand metabolism….Likewise, we demonstrated that the cellular ROSlevels in OCM/AS-MΦs derived from PBMC MΦs were significantlyupregulated, whereasthe addition of Cleanascite™ mitigated the increasedascites-mediated ROS level ,indicating that the accumulation of intercellular UFAs is responsiblefor the enhanced ROS production in OCM-MФs.” The article goes onto state “ Incontrast, removing lipids by Cleanascite™ remarkably prevented thereduction of YAP1 in OCM-MΦs .

Zhu,Meilin, et al. " VaginalLactobacillus fatty acid response mechanisms reveal ametabolite-targeted strategy for bacterial vaginosistreatment ." Cell (2024).Bacterial vaginosis (BV), a common syndrome characterizedby Lactobacillus-deficientvaginal microbiota, is associated with adverse health outcomes. BVoften recurs after standard antibiotic therapy in part becauseantibiotics promote microbiota dominance by Lactobacillusiners insteadof Lactobacilluscrispatus .Strategies to promote L.crispatus andinhibit L.iners arethus needed. This research shows that oleic acid (OA) and similarlong-chain fatty acids simultaneously inhibit L.iners andenhance L.crispatus growth.The article states “ Toprepare lipid-depleted MRS+CQ broth, four parts MRS+CQ broth wascombined with one part Cleanascite™ …. The media supernatant was collected and re-sterilized.”

Zhang,T., Zhao, F., Hu, Y. etal . Environmentalmonobutyl phthalate exposure promotes liver cancer via reprogrammedcholesterol metabolism and activation of the IRE1α-XBP1spathway. Oncogene (2024). https://doi.org/10.1038/s41388-024-03086-1

Abnormalitiesin lipid metabolism is a molecular hallmark for not only cancercells, but also tumor-associated immune cells. To study thoseabnormalities, the article states “ To investigate the role oflipids in regulating the macrophage function, wefirst employed Cleanascite to remove all lipids and lipoproteins fromthe conditioned media (CM) of HepG2 cells. Interestingly, lipid-depleted HepG2 CM significantlysuppressed the migration ability of THP-1PMA cells both in controland MBP-exposed groups. Also, the expression of M2-type markers (suchas CD206, CD163, and ARG1) in THP-1PMA cells dramatically decreasedafter lipid clearance, while the expression of M1-type markers wasup-regulated…”

Flury,Anna, et al. " Integratedstress response associated with dark microglia promotes microgliallipogenesis and contributes to neurodegeneration ." bioRxiv (2024):2024-03. Theauthors removed fatty acids, lipoproteins, lipid-membrane bodies, andother lipids from the SLB-conditioned media using Cleanascite™.

El-Hattab,Mariam Y., et al. "Nativeadiponectin plays a role in the adipocyte-mediated conversion offibroblasts to myofibroblasts ." Journalof the Royal Society interface 20.202(2023): 20230004. “wedepleted lipids (LD) and lipid-associated factors using Cleanasciteand found the α-SMA inducing activity of ACM was also eliminated…”

Gudgeon,Nancy, et al. " Uptakeof long-chain fatty acids from the bone marrow suppresses CD8+ T-cellmetabolism and function in multiple myeloma. " BloodAdvances (2023). Thearticle states “… toprobe a role for lipids in this suppressive activity of BM plasma,they were depleted from BM samples (through the use of Cleanascite™) and compared with lipid-replete BM samples.

Sinha,S., Aizawa, S., Nakano, Y. etal. Hepaticstellate cell stearoyl co-A desaturase activates leukotriene B4receptor 2 - β-catenin cascade to promote liver tumorigenesis. NatCommun 14,2651 (2023). Thearticle states “ Lipidremoval from the <10 kDafiltrate with thelipid adsorption reagent (Cleanascite ^TM)abolished the TEADpromoter stimulatory activity…”

Albakri,Marwah M., et al. "Fattyacids secreted from head and neck cancer induce M2 ”likeMacrophages ." Journalof Leukocyte Biology (2022).

Toassess depletion of fatty acids from tumor supernatants,tumor-conditioned medium was treated with Cleanascite according tothe manufacturer’s instructions and prior to incubation withmonocytes. The article states “Depletionof Fatty acids with Cleanascite from FaDu or SCC supernatants largelyreversed the phenotypic changes in Macrophage sotherwise observed by incubating monocytes in these supernatants”.

Chan,David, et al. " PolyunsaturatedFatty Acids Promote Protumoral Macrophage Polarization via aRhoA-YAP1 Signaling Pathway in the Ovarian Cancer Microenvironment ."(2022).

Thearticle states “ Toselectively remove lipids, Cleanascite™ Lipid Removal Reagent(Biotech Support Group) was added to the omental conditioned medium(OCM) according to the manufacturer’s suggestions. …Intriguingly, the removal of free fatty acids in OCM by theCleanascite™ attenuated lipid droplets deposition in M2-like MΦsand OCM-MΦs (Fig. 4A), indicating protumoral M2-like TAMs exhibited higher lipidaccumulation and metabolism in the fatty acid-enriched OCM or themalignant ascites.”

Pham,Ly, et al. " Neutrophiltrafficking to the site of infection requires Cpt1a-dependent fattyacid β-oxidation ." CommunicationsBiology 5.1(2022): 1-13. Thearticle states for lipid depletion “…cells were incubated withRPMI containing 0.5% FBS or with 10% FBS that had been treatedwith Cleanascite to remove lipids ”.

Jiang,Lifeng, et al. " Akt-DependentGlycolysis-Driven Lipogenesis Supports Proliferation and Survival ofHuman Pulmonary Arterial Smooth Muscle Cells in PulmonaryHypertension ." Frontiersin medicine 9(2022)

Thearticle states “ Toachieve the lipid-deprived cell culture condition ,cell culture grade fetal bovine serum (FBS) and BSA were delipidatedusing Cleanascite ™ …”

Yang,X. U. A. N., et al. " SCD1/FADS2fatty acid desaturases equipoise lipid metabolic activity andredox-driven ferroptosis in ascites-derived ovarian cancer cells ."(2021).

Themechanisms underlying ovarian cancer (OvCa) cells dictating theirlipid metabolic activities in promoting tumor progression remainelusive. The article states: “Comparedwith the negative controls (OCM pretreated with the lipid removalreagent, Cleanascite) ,OvCa cells cocultured in the lipid-enriched OCM showed an increase of18% in membrane fluidity.”

Pointner,Lisa, et al. " Birchpollen induces Toll-like receptor 4-dependent dendritic cellactivation favoring T cell responses. " Frontiersin Allergy (2021): 42.

Thearticle states “Toremove the lipids in birch pollen extracts, Cleanascite™ was used…according to manufacturer's recommendations … in a ratio 1:1(v/v). Importantly, “non-specifictreatment-associated and cytotoxic effects were ruled out …asneither the protein digestion nor the lipid extraction procedureaffected cell activation.”

Wang,Xueyu, et al. "EpigeneticSilencing of miR-33b Promotes Peritoneal Metastases of Ovarian Cancerby Modulating the TAK1/FASN/CPT1A/NF-κ BAxis." Cancers 13.19(2021): 4795. Theeffective use of Cleanascite™ helped establish that “… depletionof fatty acids by Cleanascite in OCM significantly impaired ovariancancer cell migration and invasion.”

Chen,Rain R., et al. " Targetingof lipid metabolism with a metabolic inhibitor cocktail eradicatesperitoneal metastases in ovarian cancer cells ." CommunicationsBiology 2(2019).

Thearticle states: ”To determine whether fatty acids in OCM are theprimary energy source, fatty acids from OCM was first removed byCleanascite™ Lipid Removal Reagent... Then, XTTcell proliferation assays showed that the growth rate of ovariancancer cells was remarkably reduced in cells cultured inCleanascite™-treated OCM. Likewise, co-treatment with Cleanascite™and OCM significantly attenuated the increased cell migration andinvasion capacities of ES-2 and SKOV3 cells .”.

Lee,Hong-Jai, et al. "Regulatoryeffect of humoral milieu on the viral DNA and surface antigenexpression of hepatitis B virus (HBV) in vitro. " Molecular& Cellular Toxicology 15.2 (2019): 123-128.Thelevels of HBsAg and HBV DNA were significantly decreased with lipidremoval by Cleanascite™ in mouse serum rather than human serum”.

Dean,E. Danielle, et al. "Interruptedglucagon signaling reveals hepatic α cell axis and role forL-glutamine in α cell proliferation. " Cellmetabolism 25.6(2017): 1362-1373. Thearticle states “For lipid removal, wholemouse serum was treated with Cleanascite™ prior to islet. Lipidremoval was validated by HPLC to remove 99% of all phopsholipids,cholesterols, and triglycerides ….”.

MonoclonalAntibodies/Ascites

Testa,Anna, et al. " TARGETINGTHE αVβ3/NgR2 PATHWAY IN NEUROENDOCRINE PROSTATE CANCER ." MatrixBiology (2023).

Thearticle states “LM609 purification was performed as previouslydescribed. Briefly, LM609 ascites was cleaned with Cleanascite …, and recirculated over a Protein A column.”

Shapiro,Scott, et al. " ImmunoglobulinG monoclonal antibodies to Cryptococcus neoformans protect micedeficient in complement component C3 ." Infectionand immunity 70.5(2002): 2598-2604.

“Theascites fluid was collected and centrifuged to remove cells. Lipidsand cell debris were removed with Cleanascite .”


TissueProteomics

Thomas,R., et al. " ToxicologicalProfile of Chemical Interference of Thyroid Hormone Metabolism viaDeiodinase Enzymes In a Rodent Model ." ToxicologicalSciences (2025):kfaf141. In part through the addition of Cleanascite™,optimization of the LC-MS/MS based assay described here permittedprecise estimates of deiodinase action relative to alternativeapproaches and offers new insights into the broader implications ofchemical interference.

TissueProteomics

Guldner,Ian H., et al. "Synapticproteins that aggregate and degrade slower with aging accumulate inmicroglia ." bioRxiv (2025):2025-05. To characterize homogenized mouse brain tissues to evaluateproteome turnover with aging, the article states “Lipidswere removed by the addition of 10 µL Cleanascite …per40 µL homogenate…”. Upon further processing, samples wereanalyzed by liquid-Chromatrography – Mass Spectrometry (LC-MS) forproteomic comparisons.

Tissue Proteomics

Guldner,Ian H., et al. "Ageingpromotes microglial accumulation of slow-degrading synapticproteins ." Nature (2026):1-12. In part through the use of Cleanascite™, combining proteinaggregate isolation techniques with neuronal BONCAT labelling helpeddefine the neuronal aggregome, a catalogue of neuronal proteins thatcontribute to protein aggregates in the aged brain.

Extracellularvesicle clearance/cell response

Nguyen,Doan C., et al. "Extracellularvesicles from bone marrow-derived mesenchymal stromal cells supportex vivo survival of human antibody secreting cells ." Journalof extracellular vesicles 7.1(2018): 1463778.

Cleanascite™-treatmentof the secretome dramatically reduced ASC functional survival, …Similar reductions were also noted with the secretome ofnon-irradiated MSC when treated with Cleanascite ™…”

daSilva Barreira, David, et al. "Membranevesicles released by Lacticaseibacillus casei BL23 inhibit thebiofilm formation of Salmonella Enteritidis ." ScientificReports 13.1. “Next,we decided toremove all lipids from LC-CFS >100and LC-CFS >3fractions and their corresponding controls (MRS >100and MRS >3)using a lipid removal reagent…Cleanascite; following the manufacturer’s instructions.” “The antibiofilmactivity of the delipidated fractions (Lipid −) was then comparedto the initial fractions (Lipid +)by crystal violet staining (Fig.) …wesaw a significant increase in biomass formation with the delipidatedLC-CFS > 3fraction compared to the control (MRS > 3)and the untreated fractions.”

Biofluids

Arora,Pearl, et al. " Nanopore”baseddetection of periodontitis biomarker miR31 in salivasamples. " Electrophoresis (2024).

Aberrationsin the microRNAs (miRNA) levels have been the cause behind variousdiseases, including periodontitis. In this study, the researchersdeveloped a label-free, real-time sensing method for the detection ofmiR31. When human saliva was tested however, extended pore blockagewith large noise levels was noted due to the presence of interferinglipid or lipoprotein complexes. To circumvent this, the articlestates “we found that that the nonspecific pore blockage by thesample matrix could be significantly diminished bysimply treating human saliva samples with Cleanascite. …By takingadvantage of Cleanascite treatment ,the saliva samples produced current modulations with similarsignatures to that of the miR31-p5 duplex in the control (i.e., inwater solutions).”

Guyon,Léna, Anne-Claire Groo, and Aure” lieMalzert-Fre ”on. "RelevantPhysicochemical Methods to Functionalize, Purify, and CharacterizeSurface-Decorated Lipid-Based Nanocarriers ." MolecularPharmaceutics (2020).

“Conjugationof PPACK (a short chain peptide that inhibits thrombin) to liposomeswas investigated through HPLC quantification of uncoupled peptiderecovered from the supernatant after centrifugation of predialysis PPACK-liposomesmixed with Cleanascite™ lipid adsorption reagent .This indirect quantification was performed at a wavelength of 215 nm(detection of amide bond).”

QinWH, Liu JT, Wang SP, Yang ZS, Wang KK, Hu B. Antibodyarray-based proteomic screening of novel biomarkers in malignantbiliary stricture .Cancer Biomark. 2022;33(3):359-368. doi: 10.3233/CBM-210095. PMID:34542063.

Thearticle states “…bile was collected upstream from the biliarystricture, then mixed with PBS containing 0.1% Tween 20 and aprotease inhibitor cocktail …,the supernatant was mixed with Cleanascite™ (Biotech Support Group,NJ, USA) to remove lipids. The article concludes that twenty proteins were found differentiallyexpressed in malignant versus benign biliary strictures…”

Plasma/Serum

Zhao, Yuan, etal. "ALabel-Free Nanopore Biosensor for Rapid and Highly SensitiveDetection of Actinomycin in Human Serum ." ACSMeasurement Science Au (2025). To overcome serum matrix effects, the samples were first treatedwith Cleanascite™,demonstrating nanopore detection that holds promise for clinicalapplications.

Playford,M. P., et al. "HDL-associatedvitamin D binding protein levels are inversely associated withnecrotic plaque burden in psoriasis ." AtherosclerosisPlus 59(2025): 32-38. Free Vitamin D Binding Protein (S-DBP) (also known asGene GC) is an abundant protein in human serum, while a smaller poolis bound to lipoproteins like HDL and VLDL. The investigators soughtto determine whether HDL-associated DBP and/or total serum DBP couldserve as biomarkers for disease severity in psoriasis andcardiovascular disease. The article states “…determine whetherDBP was found in HDL particles or just as a co fractioning proteinfrom chromatography. Fractions 20 to 28 weretreated with a lipid-removal agent (Cleanascite™) ,and while this treatment removed many of the proteins in fractions 21to 27, the non-lipidated ‘albumin’ fraction remained. At alltested concentrations of DBP, most remained unbound to the lipidremoval agent demonstrating that DBPdoes not bind to Cleanascite™ in the absence of lipid .”

HuneaultHE, Lo JS, Bai S, He Z, McPhaul MJ, Bril F, et al. Fastingintact insulin by mass spectrometry is associated with metabolicdysfunction-associated steatotic liver disease in youth .Hepatol Commun. 2024;8:e0582. https://doi.org/10.1097/ HC9.0000000000000582.Metabolic dysfunction–associated steatotic liver disease (MASLD),formerly known as NAFLD, is the most common liver disease in childrenand is a leading cause of liver-related morbidity and mortality. Thisstudy sought to determine whether fasting intact insulin, measured byliquid chromatography-tandem mass spectrometry, is associated withMASLD in children. The methods describe “Quest Diagnostics NicholsInstitute performed measurements of intact insulin and C-peptide…the samples were plated and mixed with the addition of internalstandards (bovine insulin and a stable isotopically labeled [13C/15N]C-peptide) andCleanascite delipidation reagent .”

Zheng,Wenshu, et al. " Nanopore-baseddisease diagnosis using pathogen-derived tryptic peptides fromserum ." NanoToday 45 (2022): 101515. The article states “Treatment of digestedserum samples with two commercial lipid removal agents revealedvariable effectiveness in reducing these artifacts, with one reagent(LRA; synthetic calcium silicate hydrate) revealing little to noeffect to reduce the frequency, duration, and extent of non-specificpore blockages… Serumdigests treated with the second reagent (Cleanascite™) demonstratedthe absence of these artifacts ”.

Chen,Chao, et al. " Obesity-drivenoleoylcarnitine accumulation in tumor microenvironment promotesbreast cancer metastasis-like phenotype ." ActaPharmaceutica Sinica B (2025).The article states “Forthe delipidation of mouse serum, Cleanascite was employed following the manufacturer’s protocol designedspecifically for serum samples.

Dean,E. Danielle, et al. " Interruptedglucagon signaling reveals hepatic α cell axis and role forL-glutamine in α cell proliferation. " Cellmetabolism 25.6(2017): 1362-1373. Thearticle states “For lipid removal, wholemouse serum was treated with Cleanascite™ prior to islet. Lipidremoval was validated by HPLC to remove 99% of all phopsholipids,cholesterols, and triglycerides ….”.

Taylor,Steven W., et al. " Ahigh-throughput mass spectrometry assay to simultaneously measureintact insulin and C-peptide ."Clinica Chimica Acta (2016). Cleanascite™is shown both to improve LC-MS measurements, and validated inaccordance with CLIA ’88 guidelines.

Fora complete list of all Cleanascite™ Lipid Removal references,visit: http://www.biotechsupportgroup.com/References-s/138.htm#delipidation