Fig. 6 Immunological clonal prediction status improves significantly after MAF error and bias correction. Immunological clonal prediction status improves.

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Fig. 6 Immunological clonal prediction status improves significantly after MAF error and bias correction. Immunological clonal prediction status improves significantly after MAF error and bias correction. (A and B) Comparison of the top 100 frequency-ranked clonotypes with their corresponding somatic hypermutation and intraclonotype diversity index values. Bubble size represents the median number of nonsilent nucleotide somatic hypermutations per clonotype. Uncorrected data show poor separation based on immune status (red, hyperimmunized; blue, untreated mice). MAF error and bias corrected clonotypes are clearly separated on the basis of these three parameters. (C and D) Applying a stepwise nominal logistic regression, we determined the significant model parameters that describe the separation of clonotype data based on immune status in a multivariate fashion (see Supplementary Materials and Methods). Using three combinations of four training data sets (top 100 frequency-ranked clonotypes, n = 2 untreated, and n = 2 hyperimmunized) and two test data sets (top 100 frequency-ranked clonotypes, n = 1 untreated, and n = 1 hyperimmunized), we show the combined results from the test data sets (n = 3 untreated and n = 3 hyperimmunized). The y axis represents the model prediction probability of whether a given clonotype belongs to the hyperimmunized group. The uncorrected data have a low resolving power, whereas the MAF error and bias corrected data show significant separation. (E and F) Comparison of the sensitivity and specificity of the nominal logistic regression models. The receiver operating characteristics and area under the curve (AUC) for nominal logistic regression models are shown for the significant factors using uncorrected and MAF-corrected data (for model performance using all factors, see fig. S20). The Ig-seq data sets used in this figure consisted of 1 × 106 preprocessed full-length antibody reads and were obtained from library sample preparations of splenic cDNA with synthetic spike-ins from hyperimmunized mice (n = 3) and untreated mice (n = 3) (the data sets used for hyperimmunized are IM_1a, IM_2, and IM_3; the data sets used for untreated are UM_1, UM_2, and UM_3; see table S7). Tarik A. Khan et al. Sci Adv 2016;2:e1501371 Copyright © 2016, The Authors