Characterization of ONE-specific IgM mAbs.

<p>(<b>A</b>) Cross-reactivity of the ONE-specific IgM mAbs established from the MFG-E8^−/− mice. The IgM titer of the mAbs was determined by a direct antigen ELISA using dsDNA and native and aldehyde-modified BSA as the absorbed antigens. The results represent the means ± SD of three separate experiments performed in duplicate determinations. Asterisks indicate a significant difference between BSA and ONE-modified BSA groups: *, P<0.05; **, P<0.01; ***, P<0.001. Differences were analyzed by the unpaired two-tailed Student’s t test. (<b>B</b>) Sequence alignment of the hypervariable regions of ONE-specific IgM mAbs prepared from female MFG-E8^−/− mice. CDR1, CDR2, and CDR3 represent the Kabat definition complementarity-determining regions (CDRs). Hyphens (−) indicate sequence gaps, and the dots (•) indicate the sequence not determined. Sequences were aligned using the program CLUSTALW and were manually modified. Accession numbers for the sequences are as follows. Gen Bank: 9E10, CAN87019. Protein data bank: 26-2f, 1H0D; anti-Hiv-1, 1MF2. (<b>C</b>) V region gene use of ONE-specific IgM mAbs. All sequencing analyses were performed at least three times.