Plasma Autoantibody Reactivity Profiling

Plasma autoantibody reactivity was studied using full-length human protein arrays (ProtoArray v5.1, PAH05251020, Thermo Fisher) [40 (link), 41 ]. The patient and two healthy blood donors were investigated in the same experiment. Protein arrays were probed with plasma at a dilution of 1:2000, and otherwise following the manufacturer’s protocol. Protein arrays were first incubated with blocking buffer (PA055, Life Technologies) for 1 h, followed by 90-min incubation with plasma, and lastly, a 90-min incubation with detection antibodies: Alexa Fluor 647 goat anti-human IgG antibody (A21445, Thermo Fisher) at 1:2000 dilution and Dylight 550 goat anti-GST (#DY550011-13–001, Cayman Chemicals) at 1:10,000 dilution. The Innopsys InnoScan 1100 AL 3-channel ultra-high resolution microarray scanner was used for detection.

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Abolhassani H., Landegren N., Bastard P., Materna M., Modaresi M., Du L., Aranda-Guillén M., Sardh F., Zuo F., Zhang P., Marcotte H., Marr N., Khan T., Ata M., Al-Ali F., Pescarmona R., Belot A., Béziat V., Zhang Q., Casanova J.L., Kämpe O., Zhang S.Y., Hammarström L, & Pan-Hammarström Q. (2022). Inherited IFNAR1 Deficiency in a Child with Both Critical COVID-19 Pneumonia and Multisystem Inflammatory Syndrome. Journal of Clinical Immunology, 42(3), 471-483.

Publication 2022

ProtoArray v5.1 PA055 Alexa Fluor 647 goat anti-human IgG antibody Dylight 550 goat anti-GST InnoScan 1100 AL

Alexa fluor 647 Anti igg Antibodies Antibody Autoantibody Blood donors Buffer Cayman Dilution Goat Human Microarray Patient Plasma Protein arrays

Corresponding Organization :

Other organizations : Karolinska Institutet, Uppsala University, Rockefeller University, Tehran University of Medical Sciences, Children's Medical Center, Karolinska University Hospital, École Normale Supérieure de Lyon, Centre International de Recherche en Infectiologie, Inserm, Université Claude Bernard Lyon 1

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Variable analysis

independent variables

Plasma dilution (1:2000)

dependent variables

Plasma autoantibody reactivity

control variables

Blocking buffer (PA055, Life Technologies)
Incubation time (1 h for blocking, 90 min for plasma, 90 min for detection antibodies)
Detection antibodies (Alexa Fluor 647 goat anti-human IgG antibody, Dylight 550 goat anti-GST)
Microarray scanner (Innopsys InnoScan 1100 AL 3-channel ultra-high resolution)

controls

Two healthy blood donors
Not explicitly mentioned

Annotations

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