Dataset Information

A Reference Standard for Analytical Testing of Erythropoietin.

ABSTRACT:

Purpose

Erythropoietin (EPO) is a 165 amino acid protein that promotes the proliferation of erythrocytic progenitors. A decrease in endogenous EPO production causes anemia that can be treated with recombinant Human EPO (rHuEPO).

Objective

To ensure the safety and efficacy of the rHuEPO, manufacturers must use analytical methods to demonstrate similarity across batches and between different products. To do this they need reference standards to validate their equipment and methods.

Method

We used peptide mapping, size-exclusion chromatography, glycoprofiling, and isoelectric focusing to analyze a rHuEPO reference standard.

Results

Characterization demonstrates that our rHuEPO reference standard meets the criteria for quality.

Conclusion

The rHuEPO reference standard is fit for purpose as a tool for validating system suitability and methods.

SUBMITTER: Tu H

PROVIDER: S-EPMC8986685 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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Publications

A Reference Standard for Analytical Testing of Erythropoietin.

Tu Huiping H Carrick Kevin K Potts Rebecca R Hasselberg Mark M Verdecia Mark M Burns Chris C Cowper Ben B Atouf Fouad F

Pharmaceutical research 20220315 3

<h4>Purpose</h4>Erythropoietin (EPO) is a 165 amino acid protein that promotes the proliferation of erythrocytic progenitors. A decrease in endogenous EPO production causes anemia that can be treated with recombinant Human EPO (rHuEPO).<h4>Objective</h4>To ensure the safety and efficacy of the rHuEPO, manufacturers must use analytical methods to demonstrate similarity across batches and between different products. To do this they need reference standards to validate their equipment and methods.< ...[more]

PMID: 35292912

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Project description:Abstract: BACKGROUND: Obtaining reliable and reproducible two-color microarray gene expression data is critically important for understanding the biological significance of perturbations made on a cellular system. Microarray design, RNA preparation and labeling, hybridization conditions and data acquisition and analysis are variables difficult to simultaneously control. A useful tool for monitoring and controlling intra- and inter-experimental variation is Universal Reference RNA (URR), developed with the goal of providing hybridization signal at each microarray probe location (spot). Measuring signal at each spot as the ratio of experimental RNA to reference RNA targets, rather than relying on absolute signal intensity, decreases variability by normalizing signal output in any two-color hybridization experiment. RESULTS: Human, mouse and rat URR (UHRR, UMRR and URRR, respectively) were prepared from pools of RNA derived from individual cell lines representing different tissues. A variety of microarrays were used to determine percentage of spots hybridizing with URR and producing signal above a user defined threshold (microarray coverage). Microarray coverage was consistently greater than 80% for all arrays tested. We confirmed that individual cell lines contribute their own unique set of genes to URR, arguing for a pool of RNA from several cell lines as a better configuration for URR as opposed to a single cell line source for URR. Microarray coverage comparing two separately prepared batches each of UHRR, UMRR and URRR were highly correlated (Pearson's correlation coefficients of 0.97). CONCLUSION: Results of this study demonstrate that large quantities of pooled RNA from individual cell lines are reproducibly prepared and possess diverse gene representation. This type of reference provides a standard for reducing variation in microarray experiments and allows more reliable comparison of gene expression data within and between experiments and laboratories. This SuperSeries is composed of the SubSeries listed below.

Dataset Information

A Reference Standard for Analytical Testing of Erythropoietin.

Purpose

Objective

Method

Results

Conclusion

Publications

A Reference Standard for Analytical Testing of Erythropoietin.

Similar Datasets

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