Comparison of Ovalbumin Quantification Using Forward-Phase Protein Microarrays and Suspension Arrays

Published

July 7, 2006

Author(s)

Lili Wang, Kenneth D. Cole, Hua-Jun He, Diane K. Hancock, Adolfas K. Gaigalas, Y Zong

Abstract

We employed ovalbumin (a simulant used for ricin and botulism toxins in biodefense applications) and its high affinity polyclonal antibody as a model system to examine the sensitivity, dynamic range, linearity, and reproducibility of forward-phase array results in comparison to suspension arrays. It was found that protein microarrays had a dynamic range of ~4 orders of magnitude and a sensitivity of less than 1 pg/mL, respectively. The dynamic range and sensitivity of suspension arrays were close to 2 orders of magnitude and 0.25 ng/mL, respectively. The sensitivity we observed for the suspension arrays is comparable to that reported for ELISA-type assays in the literature. We used ovalbumin samples with two different purities, 38.0% and 75.8% (w/w), as determined by polyacrylamide gel electrophoresis (PAGE). These samples were used to evaluate the effect of impure samples on detection. The data obtained from the forward-phase protein arrays gave values that were consistent with the PAGE data. The data from the suspension arrays was not as consistent and may indicate this format may not give as reliable data with impure samples. Knowledge of the advantages and disadvantages of the two proteomic methods would allow their more rational use in clinical diagnosis.

Citation

Journal of Proteome Research

Volume

5

Issue

No. 7

Pub Type

Journals

Keywords

accuracy, antibody specificity, detection sensitivity, dynamic range, forward-phase protein microarrays, linearity, ovalbumin quantification, reproducibility, suspension arrays

Citation

Wang, L. , Cole, K. , He, H. , Hancock, D. , Gaigalas, A. and Zong, Y. (2006), Comparison of Ovalbumin Quantification Using Forward-Phase Protein Microarrays and Suspension Arrays, Journal of Proteome Research (Accessed December 26, 2024)