Methods for the detection and serum depletion of porcine galectin‐3

Eliaz, Isaac; Patil, Aarti R.; Navarro-Alvarez, Nalú; Wang, Zhirui; Eliaz, Amity; Weil, Elaine; Eliaz, Isaac; Sachs, David H.; Huang, Christene A.

doi:10.1002/jca.21521

Cited by 4 publications

(8 citation statements)

References 27 publications

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“…The affinity columns contained LEAF™ purified (BioLegend, San Diego, California) monoclonal antibody M3/38 rat anti‐mouse Gal‐3 monoclonal antibody from hybridoma M3/38.1.2.8 HL.2 (ATCC ® TIB‐166™), covalently bound to hydrated cyanogen‐bromide activated Sepharose 4B (GE Healthcare Life Sciences, Pittsburgh, Pennsylvania) as previously described . For column perfusion, 60 mL matrix volume was packed into a GE XK50/20 column (GE Healthcare Life Sciences, Pittsburgh, Pennsylvania) with a diameter of 50 mm and bed height of ∼30 mm.…”

Section: Methodsmentioning

confidence: 99%

“…During apheresis, the inlet flow rate typically ranged between 12–14 mL/minute resulting in a column retention time of ∼4–5 minutes, calculated based on the column having an internal diameter of 50 mm and the Sepharose matrix bed height being approximately 3 cm. To assess the binding efficiency of porcine Gal‐3 to the M3/38 antibody, plasma samples from treated animals were taken pre‐ and post‐passage through the affinity column during apheresis, and Gal‐3 concentrations quantified by sandwich enzyme‐linked immunosorbent assay (ELISA) as previously described . The Gal‐3 specific ELISA has a linear range from 0.3 to 20 ng/mL with a lower limit of detection of 300 pg/mL.…”

Section: Methodsmentioning

confidence: 99%

“…To assess the binding efficiency of porcine Gal-3 to the M3/38 antibody, plasma samples from treated animals were taken pre-and post-passage through the affinity column during apheresis, and Gal-3 concentrations quantified by sandwich enzyme-linked immunosorbent assay (ELISA) as previously described. 21 The Gal-3 specific ELISA has a linear range from 0.3 to 20 ng/mL with a lower limit of detection of 300 pg/mL. Gal-3 levels in sera from systemic blood drawn before and after the apheresis procedure were also measured.…”

Section: Apheresis Column Preparation and Perfusionmentioning

confidence: 99%

“…Given its capacity to bind β‐galactosides, we previously demonstrated that it is possible to remove Gal‐3 in vitro from plasma and serum samples using affinity chromatography mini‐columns with different substrates with greater selective affinity to Gal‐3 . Sepharose coupled with a monoclonal antibody that binds Gal‐3 could remove Gal‐3 from porcine sera ex vivo.…”

Section: Introductionmentioning

confidence: 99%

“…Given its capacity to bind β-galactosides, we previously demonstrated that it is possible to remove Gal-3 in vitro from plasma and serum samples using affinity chromatography mini-columns with different substrates with greater selective affinity to Gal-3. 21 Sepharose coupled with a monoclonal antibody that binds Gal-3 could remove Gal-3 from porcine sera ex vivo . Here, we describe an apheresis column perfusion system using affinity chromatography with Sepharose-bound to the Gal-3 monoclonal antibody and demonstrate its use in a porcine model of skin inflammation using Complete Freund’s Adjuvant (CFA) injection.…”

Section: Introductionmentioning

confidence: 99%

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The effects of galectin‐3 depletion apheresis on induced skin inflammation in a porcine model

Navarro-Alvarez

Gonçalves

Andrews

et al. 2018

J of Clinical Apheresis

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Galectin-3 (Gal-3), a β-galactoside-binding lectin that is expressed in mammalian cells, is known to modulate several biological functions such as cell-cell adhesion, macrophage activation, angiogenesis, metastasis, and fibrosis. The goal of this study was to evaluate the ability of Gal-3 depletion apheresis using an adsorption column with immobilized anti-Gal-3-antibody to reduce inflammation induced by Complete Freund's Adjuvant injection in a skin inflammation porcine model. Here, we report that plasma perfusion by apheresis through a Gal-3 binding immuno-affinity column reduces plasma Gal-3 levels to below limits of quantitative detection, and results in significant decrease in skin inflammation, including degree and duration of inflammatory lesions. Human plasma was tested ex vivo and found to be efficiently depleted using the anti-Gal-3 affinity column. This study demonstrates the potential of Gal-3 depletion apheresis as a therapeutic method for inflammation-mediated disease, supporting continued research in this area for clinical application.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Apheresis Column Preparation and Perfusionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The effects of galectin‐3 depletion apheresis on induced skin inflammation in a porcine model

Navarro-Alvarez

Gonçalves

Andrews

et al. 2018

J of Clinical Apheresis

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Multimeric fusion single‐chain variable fragments as potential novel high‐capacity ligands

et al. 2020

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In basic and applied biotechnology, design of affinity ligands has become essential for high‐capacity applications such as affinity‐based downstream processes for therapeutic molecules. Here, we established a proof‐of‐concept for the use of multimeric fusion single‐chain variable fragment (scFvs) as high‐capacity ligands in affinity adsorbents. Mono‐ and di/tri‐scFvs separated by Pro‐rich negatively charged linkers were designed, produced, and immobilized to 6% cross‐linked agarose beads. Frontal binding experiments with a target protein of 50 kDa resulted in up to 20 mg·mL−1 and 82% in dynamic binding capacity and utilization yield, respectively, at 100% breakthrough. The utilization of the binding sites was impacted by the ligand format and ligand density, rather than limitation in pore size of adsorbent as previously suggested. Overall, we demonstrated that multimeric fusion scFvs can successfully be developed and used as high‐capacity ligands in affinity adsorbents, enabling lean process design and alignment with process specifications.

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Easily Multiplexable Immunoplatform to Assist Heart Failure Diagnosis through Amperometric Determination of Galectin‐3

et al. 2020

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This work reports the first electrochemical immunoassay involving magnetic microbeads (MBs) for the determination of galectin-3 (Gal-3), a β-galactosidasebinding lectin that acts as mediator of heart failure (HF). MBs-captured sandwich-type immune complexes and amperometric detection at disposable screen-printed carbon electrodes were used. The immunoplatform showed a detection limit of 8.3 pg mL À 1 , good reproducibility, and excellent selectivity. The endogenous concentration of Gal-3 in human plasma from HF patients was determined with results in agreement with those obtained using ELISA. The multiplexing feasibility of the developed immunoplatform was demonstrated for the simultaneous determination of Gal-3 and N-terminal pro-brain natriuretic peptide (NT-proBNP).

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Methods for the detection and serum depletion of porcine galectin‐3

Abstract: Methods have been developed for the detection and depletion of porcine Gal-3. These methods will be used to study the specific effects of Gal-3 depletion via apheresis in porcine models of disease.

Cited by 4 publications

References 27 publications

The effects of galectin‐3 depletion apheresis on induced skin inflammation in a porcine model

The effects of galectin‐3 depletion apheresis on induced skin inflammation in a porcine model

Multimeric fusion single‐chain variable fragments as potential novel high‐capacity ligands

Easily Multiplexable Immunoplatform to Assist Heart Failure Diagnosis through Amperometric Determination of Galectin‐3

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