quantikine elisa mouse erythropoietin immune assay 154 kit  (R&D Systems)

Journal: Nucleic Acids Research

Article Title: RNA encoded peptide barcodes enable efficient in vivo screening of RNA delivery systems

doi: 10.1093/nar/gkae648

Figure Lengend Snippet: PALS is a good surrogate for ELISA-based protein quantitation. ( A ) LNP particle characteristics including size, polydispersity index (PDI) and encapsulating efficiency. Values are mean ± SD, n = 5. ( B ) Quantitation of bhEPO amounts in mouse serum 6 h after LNP administration in vivo . Values are mean ± SD, n = 5. ( C ) quantitation of bhEPO using barcodes after single LNP administration and using PALS. No difference in ranking of LNPS based on barcode amounts between both methods (Wilcoxon matched pairs signed rank test, P = 0.0625). ( D ) Correlation between EPO protein quantification by ELISA and by mass spectrometry using one representative EPO peptide. ( E ) comparison of protein quantitation using ELISA and PALS. No difference in normalized mean between both methods (paired t -test) and no difference in ranking of LNPs using both methods (Wilcoxon matched-pairs signed rank test, P = 0.3750). ( F ) correlation of protein quantitation by ELISA and PALS using the peptide barcodes.

Article Snippet: Human EPO levels in cell culture medium and from mouse serum samples was analysed using a human EPO Quantikine IVD ELISA kit (DEP00, R&D Systems, Abingdon, UK) following the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Protein Quantitation, Quantitation Assay, In Vivo, Mass Spectrometry, Comparison

Journal: Nucleic Acids Research

Article Title: RNA encoded peptide barcodes enable efficient in vivo screening of RNA delivery systems

doi: 10.1093/nar/gkae648

Figure Lengend Snippet: PALS is sensitive to differentiate LNPs and rank them based on functionality. ( A ) LNP particle characteristics including size, polydispersity index (PDI) and encapsulating efficiency. Values are mean ± SD. ( B ) representation of bLNP combinations in each pool. Total mRNA dose per pool was 10 μg. ( C ) Quantification of EPO protein from mouse serum 6 h after i.v. administration measured by ELISA and mass spectrometry normalized to the standard LNP control. Values are mean ± SD, n = 5. ( D ) Quantification of hEPO in the individual LNP treated groups is consistent with quantification using the barcodes with a good correlation between both methods. Values are mean ± SD, n = 5. ( E ) Quantification of barcodes in the animals which received a single LNP is consistent with quantification of barcodes in animals which received the pool. Values are mean ± SD, n = 5. ( F ) Dose-response for each bLNP showing PALS is sensitive to differentiate and rank LNPs based on functionality at different dose levels. Values are mean ± SD, n = 5.

Article Snippet: Human EPO levels in cell culture medium and from mouse serum samples was analysed using a human EPO Quantikine IVD ELISA kit (DEP00, R&D Systems, Abingdon, UK) following the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Mass Spectrometry, Control

Journal: Nucleic Acids Research

Article Title: RNA encoded peptide barcodes enable efficient in vivo screening of RNA delivery systems

doi: 10.1093/nar/gkae648

Figure Lengend Snippet: PALS is a good surrogate for ELISA-based protein quantitation. ( A ) LNP particle characteristics including size, polydispersity index (PDI) and encapsulating efficiency. Values are mean ± SD, n = 5. ( B ) Quantitation of bhEPO amounts in mouse serum 6 h after LNP administration in vivo . Values are mean ± SD, n = 5. ( C ) quantitation of bhEPO using barcodes after single LNP administration and using PALS. No difference in ranking of LNPS based on barcode amounts between both methods (Wilcoxon matched pairs signed rank test, P = 0.0625). ( D ) Correlation between EPO protein quantification by ELISA and by mass spectrometry using one representative EPO peptide. ( E ) comparison of protein quantitation using ELISA and PALS. No difference in normalized mean between both methods (paired t -test) and no difference in ranking of LNPs using both methods (Wilcoxon matched-pairs signed rank test, P = 0.3750). ( F ) correlation of protein quantitation by ELISA and PALS using the peptide barcodes.

Article Snippet: Purified EPO protein was quantified by ELISA (DEP00, R&D Systems, UK) and trypsin digested using an in-solution tryptic digestion kit (89895, ThermoFisher Scientific, UK) following the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Protein Quantitation, Quantitation Assay, In Vivo, Mass Spectrometry, Comparison

Journal: Nucleic Acids Research

Article Title: RNA encoded peptide barcodes enable efficient in vivo screening of RNA delivery systems

doi: 10.1093/nar/gkae648

Figure Lengend Snippet: PALS is sensitive to differentiate LNPs and rank them based on functionality. ( A ) LNP particle characteristics including size, polydispersity index (PDI) and encapsulating efficiency. Values are mean ± SD. ( B ) representation of bLNP combinations in each pool. Total mRNA dose per pool was 10 μg. ( C ) Quantification of EPO protein from mouse serum 6 h after i.v. administration measured by ELISA and mass spectrometry normalized to the standard LNP control. Values are mean ± SD, n = 5. ( D ) Quantification of hEPO in the individual LNP treated groups is consistent with quantification using the barcodes with a good correlation between both methods. Values are mean ± SD, n = 5. ( E ) Quantification of barcodes in the animals which received a single LNP is consistent with quantification of barcodes in animals which received the pool. Values are mean ± SD, n = 5. ( F ) Dose-response for each bLNP showing PALS is sensitive to differentiate and rank LNPs based on functionality at different dose levels. Values are mean ± SD, n = 5.

Article Snippet: Purified EPO protein was quantified by ELISA (DEP00, R&D Systems, UK) and trypsin digested using an in-solution tryptic digestion kit (89895, ThermoFisher Scientific, UK) following the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Mass Spectrometry, Control

Journal: Frontiers in Physiology

Article Title: Cardiomyocyte crosstalk with endothelium modulates cardiac structure, function, and ischemia-reperfusion injury susceptibility through erythropoietin

doi: 10.3389/fphys.2024.1397049

Figure Lengend Snippet:

Article Snippet: EPO protein concentration was quantified from serum using a Quantikine Mouse EPO ELISA (MEP00B, R&D Systems) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Recombinant, Standard Deviation, Nucleic Acid Electrophoresis, Microarray