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. 2021 Dec 15;12(12):1131–1143.e5. doi: 10.1016/j.cels.2021.08.008

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© 2021 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Monitoring personalized plasma Fab repertoires reveals not only their simplicity and extreme donor uniqueness but also longitudinal clonal variations within a single donor

(A) Heatmap illustrating the degree of overlap between the detected IgG1 repertoires in all analyzed sepsis patient plasma samples. For each of the eight donors, four sampling times were available, and Fab profiles were measured by LC-MS analysis. Each LC-MS peak, exhibiting a unique mass and retention time pair, was considered a unique clone and annotated as ^RT # _mass. The amount of Fab molecules, based on intensity that is persistent, is quantified and shown as a percentage as indicated by the color bar. In between donors, the overlap is found to be on average 3%, whereas within a single donor at different time points the overlap was found to be in between 26% and 98%.

(B) Hierarchical clustering of the Fab clonal repertoires based on correlation distance. The branch lengths depict the distance between the repertoires. Donors are colored as in Figure 1A.

(C) Longitudinal deconvoluted Fab mass profiles of donor F76 at each of the four time points. Each peak represents a unique Fab at its detected mass and plasma concentration. The top 30 most intense Fab clones in each sample are colored reflecting the time points, the other clones are colored gray. Five peaks are highlighted with a box that is colored based on the longitudinal behavior of the Fab concentration in plasma (blue, diminishing clone; green, persistent clone; and red, post-sepsis clone), a magnified version of each of these Fab signals is shown in (E).

(D) Pie charts portraying the total number and distribution of clones in donor F76 for each time point. The value within the chart displays the number of identified unique Fab molecules. The five most intense Fabs are colored based on longitudinal behavior, and their mass and retention time are depicted in the legend in order of abundance.

(E) Magnified mass plots for each of the highlighted clones. The peaks are colored according to the time points, the surrounding border and sign indicate the longitudinal behavior and the top right shows the annotated clone ID.