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. 2023 Jan 6;13:306. doi: 10.1038/s41598-022-27281-w

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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SP-IRIS-based antibody competition assay for anti-EBOV GP antibodies. Top part shows a schematic of the SP-IRIS competition assay. First, antibody 1 (Ab 1) is added to rVSV-EBOV sample in the microcentrifuge tube and incubated for 20 min. Then, this mixture is applied to the SP-IRIS chip spotted with anti-EBOV GP mAbs (Ab 2) and incubated for 1 h. Each mAb is spotted with 4 replicate spots. Following incubation, binding on each mAb is calculated as a percentage of the uncompeted binding from a reference chip that was incubated with the rVSV-EBOV sample alone. Signal on the Ab 2 on the reference chip is taken as 100%. Bottom table summarizes the percent competition binding values. Ab 1 competes with Ab 2 when the percentage is lower than 50% (black boxes). When the percent binding is between 50 and 75%, it is considered intermediate competition (dark gray). Percent binding values between 75 and 110% represent no competition (white boxes), whereas the values over 110% represent an effect that we refer to as binding enhancement (light gray boxes). The results presented in Fig. 7 are representative of two independent competition assays.