A recombinant mouse interleukin-4 (IL-4) and three different purified rat anti-mouse IL-4 monoclonal antibodies (Mab) with different epitopes were employed as a model system. This system was used to examine monoclonal antibody effectiveness using both conventional and high-throughput measurement techniques in order to select epitopes for attaining the most sensitive detection of the recombinant IL-4 through the sandwich-type immunoassays. A surface plasmon resonance (SPR) measurement technique and two high-throughput methods, suspension arrays (also called multiplexed bead arrays) and forward-phase protein microarrays, predicted the same capture (BVD4-1D11) and detection (BVD6-24G2) antibody pair for most sensitive detection of the recombinant cytokine. By using this antibody pair, we were able to detect as low as 2 pg/mL of IL-4 in buffer solution and 13.5 pg/mL of IL-4 spiked in 100% of normal mouse serum with the multiplexed bead arrays. Due to large amount of material required for SPR measurements, the study suggests that the multiplexed bead arrays and protein microarrays are both suited for epitope selection of numerous antibodies against the same analyte of interest to meet the need in the era of systems biology and reproducible clinical diagnostic for better patient care.
Citation: Journal of Proteome Research
Pub Type: Journals
detection sensitivity, epitope selection, equilibrium binding constant, forward-phase protein microarrays, interleukin-4 quantification, monoclonal antibody, surface plasmon resonance, suspension arrays