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Aim. To analyze the levels of B-cell-produced antibodies in the vitreous humor of patients with or without diabetic retinopathy (DR) both qualitatively and quantitatively. Methods. A total of 52 type 2 diabetes mellitus (T2DM) with DR patients and 52 control subjects without diabetes mellitus or inflammatory diseases were included in this prospective study. The levels of immunoglobulin (Ig)A, IgM, and IgG subtypes were measured using a magnetic color-bead-based multiplex assay. Results. The concentrations of IgA, IgM, and total antibodies in the DR group were significantly higher than those in the control group (all p < 0.001), but there was no significant difference in the 4 IgG subtypes between the two groups after Bonferroni correction. Pearson's correlation analysis revealed low negative correlations between levels of antibodies (IgA, IgM) and estimated glomerular filtration rate (eGFR, r = − 0.443 , r = − 0.377 , respectively, both p < 0.05). Furthermore, multiple linear regression analysis yielded three equations to predict the concentrations of IgA, IgM, and total antibodies in the vitreous humor according to eGFR and other clinical variables (r = 0.542 , r = 0.461 , and r = 0.312 , respectively, all p < 0.05). Conclusion. Increased levels of IgA, IgM, and total antibodies produced by B cells were observed in the vitreous humor of T2DM patients with DR. There were low negative correlations between levels of antibodies (IgA, IgM) and eGFR. [ABSTRACT FROM AUTHOR]