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. 2017 Jun 26;7:4230. doi: 10.1038/s41598-017-04275-7

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© The Author(s) 2017

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Anti-LTF Abs in RA sera. Serum samples from healthy adults (NHS, n = 35) and patients with RA (n = 80) were individually 1/200 diluted and assayed in huLTF-based ELISAs for detection of huLTF-specific IgM (A) and IgG (B) Abs. The detection Abs were HRP-conjugated goat-anti-human IgM or IgG with OPD as substrate. Values are the mean OD492 nm ± SEM from triplicate wells. Horizontal lines represent group averages. NS: not statistically significant. RA sera with highest levels of anti-LTF IgG were selected and pooled for affinity purification of human anti-LTF IgG autoantibodies using Protein A- and huLTF-Sepharose 4B columns. The resultant huLTF-specific IgG (RA-IgG) was compared with the sample of flow through and also IVIG (commercial) in huLTF-based ELISAs, **p < 0.01 compared with IVIG control (C). Samples of RA sera, RA-IgG and IVIG were also analyzed by Coomassie blue-stained SDS-PAGE 10% gel (D).