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Polyreactivity ELISAs.

JG Jenna J. Guthmiller
HU Henry A. Utset
CH Carole Henry
LL Lei Li
NZ Nai-Ying Zheng
WS Weina Sun
MV Marcos Costa Vieira
SZ Seth Zost
MH Min Huang
SH Scott E. Hensley
SC Sarah Cobey
PP Peter Palese
PW Patrick C. Wilson
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Polyreactivity was determined using a polyreactive ELISA protocol, as previously described (13). Briefly, MAbs were tested for binding to 6 antigens (cardiolipin, dsDNA, flagellin, insulin, KLH, and LPS) starting at 1 μg/ml for 1.5 h at 37°C. HRP-conjugated goat anti-human IgG antibody (Jackson ImmunoResearch) diluted 1:2,000 in PBS/0.05% Tween/0.1 mM EDTA was used to detect binding of MAbs, and plates were subsequently developed with super aquablue ELISA substrate (eBiosciences). MAbs were considered polyreactive if they bound 4 or more antigens at an OD of 450 nm (OD450) of ≥0.5. All MAbs were tested in duplicate, and all assays were performed 2 or 3 times.

Copyright and License information:  ©2021 Guthmiller et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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