Comparison of ELISA with electro-chemiluminescence technology for the qualitative and quantitative assessment of serological responses to vaccination

Abstract

Background: Profiling immune responses induced by either infection or vaccination can provide insight into identification of correlates of protection. Furthermore, profiling of serological responses can be used to identify biomarkers indicative of exposure to pathogens. Conducting such immune surveillance requires readout methods that are highthroughput, robust, and require small sample volumes. While the enzyme-linked immunosorbent assay (ELISA) is the classical readout method for assessing serological responses, the advent of multiplex assays has significantly increased the throughput and capacity for immunoprofiling. This report describes the development and assay performance (sensitivity, linearity of detection, requirement for multiple dilutions for each sample, intra-and inter-assay variability) of an electro-chemiluminescence (ECLIA)-based multiplex assay. Methods:The current study describes the development of a multiplex ECLIA-based assay and characterizes the sensitivity, linear range, and inter-and intra-assay variability of the ECLIA platform and its agreement with the traditional ELISA. Special emphasis was placed on potential antigenic competition when testing closely related antigens in the multiplex format. Results:Multiplexing of antigens in ECLIA provides significant practical benefits in terms of reducing sample volume requirements and experimental time. Beyond the practical advantages of multiplexing, the ECLIA provides superior assay performance when compared to the ELISA. Not only does ECLIA show good agreement with the ELISA assay, but the linear range of ECLIA is also sufficiently wide to permit single-dilution measurements of concentration without the need to do serial dilutions. The lack of antigenic competition allows the simultaneous testing of closely related antigens, such as plate antigens representing different alleles of the same protein, which can inform about cross-reactivities-or lack thereof-of serological responses.

Serological profiles of pan-coronavirus-specific responses in COVID-19 patients using a multiplexed electro-chemiluminescence-based testing platform

“…The ECLIA-based MSD platform was chosen based on its superiority in previous studies using malarial antigens [17, 18]. In the present study, we evaluated a pan-CoV panel of recombinant proteins generated in a mammalian expression system to ensure proper glycosylation.…”

“…Although numerous SARS-CoV-2-specific serological assays have been developed [14, 15], additional challenges remain in using these assays for surveillance or clinical management [16] including: 1) assay throughput, 2) the specificity of assay readouts to SARS-CoV-2 as compared to other related coronaviruses 3) and the need to perform sample testing at multiple dilutions due to the narrow linear range of the respective assay platform. Recently, we compared a newly developed multiplexed serological assay based on electro-chemiluminescence (ECLIA) customized for malaria serosurveillance with a qualified standard ELISA [17]. The results demonstrated superiority of the ECLIA-based assay in several aspects including a wide linear range eliminating the need for serial dilutions of test samples, low variability, robust reproducibility of the assay, and no signal reduction due to antigenic competition when testing closely related antigens.…”

Breadth of humoral immune responses to the C-terminus of the circumsporozoite protein is associated with protective efficacy induced by the RTS,S malaria vaccine

“…The experiments were conducted using the U-PLEX assay platform (Mesoscale Discovery (MSD) Inc, Gaithersburg, MD) as previously reported [14]. In brief, individual biotinylated peptides were incubated with the proprietary U-PLEX MSD linkers designed to bind to respective spots on 10-spot MSD multi-array 96 plates.…”

Performance characteristics of the Mindray chemiluminescence anti‐Müllerian hormone assay

“…As compared with the traditional ELISA method, the chemiluminescence method has high sensitivity, good repeatability, a wide linear range, and high degrees of automation and standardization, and thus has been adopted in many hospitals. 11,12 About 30 hospitals in Guangdong province, China, employ the Roche AMH kit. However, the reference interval is derived from a European population.…”