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Evaluation of Immunostimulatory Potential of Branded and US-Generic Enoxaparins in an In Vitro Human Immune System Model.

Luna E, Agrawal P, Mehta R, Vernhes C, Viskov C, Amiral J, Warren WL, Drake DR - Clin. Appl. Thromb. Hemost. (2014)

Bottom Line: Low-molecular-weight heparins (LMWHs) have several positive therapeutic effects and can also form immunostimulatory complexes with plasma proteins, such as platelet factor 4 (PF4).Production of tissue factor pathway inhibitor (TFPI), a physiologic heparin-induced inhibitor of tissue factor-induced coagulation that was used as a functional readout of biological activity of enoxaparins in these assays, was heightened in the presence of branded enoxaparin complexes, but its levels were variable in cultures treated with complexes containing US-generic enoxaparins.Analytical analyses suggest that the heightened immunostimulatory potential of some of the US-generic enoxaparin product lots could be tied to their capacity to form ultra-large and/or more stable complexes with PF4 than the other LMWHs included in this study.

View Article: PubMed Central - PubMed

Affiliation: Sanofi Pasteur, VaxDesign Campus, Orlando, FL, USA.

No MeSH data available.


Differential SE-FPLC profiles of PF4–enoxaparin complexes prepared with 2 distinct lots of Amphastar. SE-FPLC analysis of Lot 1, Batch 1 and Lot 2, and Batch 1 Amphastar complexes was performed at 0.75 mol/L NaCl. PF4, platelet factor 4; SE-FPLC, size exclusion-fast performance liquid chromatography.
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fig7-1076029614562037: Differential SE-FPLC profiles of PF4–enoxaparin complexes prepared with 2 distinct lots of Amphastar. SE-FPLC analysis of Lot 1, Batch 1 and Lot 2, and Batch 1 Amphastar complexes was performed at 0.75 mol/L NaCl. PF4, platelet factor 4; SE-FPLC, size exclusion-fast performance liquid chromatography.

Mentions: Given that the MIMIC® PTE evaluations showed the 2 lots of US-generic enoxaparins had differential capacities to trigger immune activation and modulate TFPI production, we next evaluated whether the 2 lots of US-generic products would also generate unique SE-FPLC profiles under the dissociating high salt condition. As can be seen in Figure 7, the SE-FPLC profile of the complexes prepared with Lot 1, Batch 1 Amphastar generated a minor peak at the elution position of 10 on the graph (similar to Figure 6A, bottom row), whereas Lot 2, Batch 1 Amphastar generated a major peak at the graph elution position of 10. These SE-FPLC observations suggest the 2 lots of Amphastar generated PF4 complexes with unique physicochemical profiles; coupled with the results of Figure 5, it appears tightly associated complexes (yielding smaller PF4 peaks) are more immunostimulatory than more easily dissociated complexes that are capable of generating larger peaks at elution point 10.


Evaluation of Immunostimulatory Potential of Branded and US-Generic Enoxaparins in an In Vitro Human Immune System Model.

Luna E, Agrawal P, Mehta R, Vernhes C, Viskov C, Amiral J, Warren WL, Drake DR - Clin. Appl. Thromb. Hemost. (2014)

Differential SE-FPLC profiles of PF4–enoxaparin complexes prepared with 2 distinct lots of Amphastar. SE-FPLC analysis of Lot 1, Batch 1 and Lot 2, and Batch 1 Amphastar complexes was performed at 0.75 mol/L NaCl. PF4, platelet factor 4; SE-FPLC, size exclusion-fast performance liquid chromatography.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC4401814&req=5

fig7-1076029614562037: Differential SE-FPLC profiles of PF4–enoxaparin complexes prepared with 2 distinct lots of Amphastar. SE-FPLC analysis of Lot 1, Batch 1 and Lot 2, and Batch 1 Amphastar complexes was performed at 0.75 mol/L NaCl. PF4, platelet factor 4; SE-FPLC, size exclusion-fast performance liquid chromatography.
Mentions: Given that the MIMIC® PTE evaluations showed the 2 lots of US-generic enoxaparins had differential capacities to trigger immune activation and modulate TFPI production, we next evaluated whether the 2 lots of US-generic products would also generate unique SE-FPLC profiles under the dissociating high salt condition. As can be seen in Figure 7, the SE-FPLC profile of the complexes prepared with Lot 1, Batch 1 Amphastar generated a minor peak at the elution position of 10 on the graph (similar to Figure 6A, bottom row), whereas Lot 2, Batch 1 Amphastar generated a major peak at the graph elution position of 10. These SE-FPLC observations suggest the 2 lots of Amphastar generated PF4 complexes with unique physicochemical profiles; coupled with the results of Figure 5, it appears tightly associated complexes (yielding smaller PF4 peaks) are more immunostimulatory than more easily dissociated complexes that are capable of generating larger peaks at elution point 10.

Bottom Line: Low-molecular-weight heparins (LMWHs) have several positive therapeutic effects and can also form immunostimulatory complexes with plasma proteins, such as platelet factor 4 (PF4).Production of tissue factor pathway inhibitor (TFPI), a physiologic heparin-induced inhibitor of tissue factor-induced coagulation that was used as a functional readout of biological activity of enoxaparins in these assays, was heightened in the presence of branded enoxaparin complexes, but its levels were variable in cultures treated with complexes containing US-generic enoxaparins.Analytical analyses suggest that the heightened immunostimulatory potential of some of the US-generic enoxaparin product lots could be tied to their capacity to form ultra-large and/or more stable complexes with PF4 than the other LMWHs included in this study.

View Article: PubMed Central - PubMed

Affiliation: Sanofi Pasteur, VaxDesign Campus, Orlando, FL, USA.

No MeSH data available.