A mouse IgG primer set from Novagen (no. 69831-3) was used to amplify the IgG variable region on antibody heavy chain (VH) and variable region on antibody light chain (VL) regions from selected hybridoma clones. Briefly, 1 × 106 cells were collected for each cell line. Total RNA was extracted using TRIzol reagent (Thermo Fisher, no. 15596026). The first-strand complementary DNA was amplified using PrimeScript reverse transcription PCR kit from Takara (no. RR104A). PCR products with the expected size [an average size of about 400 base pairs (bp) for VH, and 360 bp for VL] were sequenced. The sequences of the PCR products were analyzed by IMGT/V-QUEST (www.imgt.org) (47) to define the VH or VL regions and the corresponding subelements. The uniqueness of antibody sequences was evaluated by comparing full-length V (VH and VL), frame, or CDR sequences using clustal algorithm. The homology matrixes were shown in the heat map format and that of the combined CDR sequences is shown in fig. S1C as an example.

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