Images of antigen microarrays and determination of linearity of the array assay.

Ferreira, Victor H.; Chruscinski, Andrzej; Kulasingam, Vathany; Pugh, Trevor J.; Dus, Tamara; Wouters, Brad; Oza, Amit; Ierullo, Matthew; Ku, Terrance; Majchrzak-Kita, Beata; Humar, Sonika T.; Bahinskaya, Ilona; Pinzon, Natalia; Zhang, Jianhua; Heisler, Lawrence E.; Krzyzanowski, Paul M.; Lam, Bernard; Lungu, Ilinca M.; Manase, Dorin; Pace, Krista M.; Mashouri, Pouria; Brudno, Michael; Garrels, Michael; Mazzulli, Tony; Cybulsky, Myron; Humar, Atul; Kumar, Deepali

doi:10.1371/journal.pone.0247258.s002

pone.0247258.s002.pdf (11.31 MB)

Images of antigen microarrays and determination of linearity of the array assay.

journal contribution

posted on 2021-02-16, 18:41 authored by Victor H. Ferreira, Andrzej Chruscinski, Vathany Kulasingam, Trevor J. Pugh, Tamara Dus, Brad Wouters, Amit Oza, Matthew Ierullo, Terrance Ku, Beata Majchrzak-Kita, Sonika T. Humar, Ilona Bahinskaya, Natalia Pinzon, Jianhua Zhang, Lawrence E. Heisler, Paul M. Krzyzanowski, Bernard Lam, Ilinca M. Lungu, Dorin Manase, Krista M. Pace, Pouria Mashouri, Michael Brudno, Michael Garrels, Tony Mazzulli, Myron Cybulsky, Atul Humar, Deepali Kumar

A) Images of 2-color arrays probed with secondary antibodies only, pre-COVID serum (negative control) and COVID+ serum (positive control). Antigens were spotted in triplicate; green indicates IgG reactivity, whereas red indicates IgM reactivity. On the array probed only with secondary antibodies, only human IgG and human IgM are detected. On the array probed with pre-COVID serum, reactivity against common community coronavirus antigens is detected. On the array probed with COVID+ serum, there are additional SARS-CoV-2 reactivities detected (boxes). Array features are approximately 500 μm in diameter. B) and C) Linearity studies using serial dilutions of COVID+ serum. Graph B shows MFI-B plotted against serum dilutions, whereas Graph C shows log2 transformed MFI-B. Linear responses are observed over a wide range of serum dilutions using log2 transformed MFI-B. Antibody responses become non-linear as MFI-B approaches saturation levels (MFI-B > 60,000). Abbreviations: MFI-B—median fluorescent intensity minus background.

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