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. 2020 Sep 23;18:262. doi: 10.1186/s12916-020-01721-8

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© The Author(s) 2020

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 5 — Characteristics of anti-Neu5Gc IgG and Neu5Gc in food. a Ratio between sum anti-Neu5Gc IgG against α3-linked-/α6-linked-Neu5Gc-glycans calculated in men 45–60 stratified based on meat Neu5Gc intake (Fig. S3). The α3/α6 linkage ratio in Q1 (n = 10) is lower than that in Q2–Q4 (n = 30) (Mann-Whitney test; p = 0.08; Q1 0.6080 ± 0.2806, Q2–Q4 1.472 ± 0.3722; mean ± sem). Similar comparing Q1 and Q4 (p = 0.09; Q1 0.6080 ± 0.2806, Q4 1.384 ± 0.3961). b ROC curve of α3/α6 linkage ratio score in Q1 (n = 10) compared to Q2–Q4 (n = 30) showed AUC 0.687 ± 0.098 (p = 0.08). Similar between Q1 and Q4 (p = 0.08, AUC 0.73 ± 0.1162). c Anti-Neu5Gc antibodies were affinity-purified from pooled sera of men 45–60 consuming low/high Neu5Gc from meat (Q1 5.39 ml and Q4 6.8 ml sera; n = 10 per group). Antibody yield was higher in Q4 than Q1 (8.01 versus 4.01 μg/ml serum, respectively). d, e IgG reactivity on sialoglycan microarrays (2 μg/block; detected with Cy3-anti-human IgG) revealed high specificity against Neu5Gc-glycans, no reactivity against Neu5Ac-glycans (d; each dot is IgG response/glycan), with altered diversity of glycans in Q4 over Q1 (e; Pearson r = 0.22). The α3/α6 linkage ratios: Q1 1.34, Q4 2.04. f Affinity (K_D) per glycan calculated from anti-Neu5Gc IgG on microarrays at 16 serial dilutions (40–4.9 × 10⁻³ ng/μl; 266.7–0.033 nM; non-linear fit with one-site specific binding), showing no change in affinities with higher Neu5Gc intake (mean ± sem; t test). g Gram of food to consume to reach daily nmol Neu5Gc per quartile based on Neu5Gc content (measured by DMB-HPLC). Q1 max value based on gr food to reach men Q1 average of 9443 nmol/day. Q4 min value is based on gr food to reach women Q4 average of 19,627 nmol/day. Gcemic index is the Neu5Gc content (nmol/gr) in each food item relative to the amount measured in beef (163 nmol/gr)