MAbs were developed using a large-scale mAb development operation modeled after an assembly line. In the antibody assembly line, each of close to 100 highly trained technicians performs one to three discrete steps (for example, plating fusion cells onto 96-well plates or cell transfer from 96- to 384-well plates) for making hybridomas. An internally built informatics and data system (Antibody Assembler) is used for tracking materials and project status. More than 90% of all materials used are bar-coded to minimize hand labeling. Many steps have automatic data analysis and decision making (for example, clone picking). Together, antibody assembly line is scalable and cost efficient. PETAL is a premade library built by this highly efficient process. After traditional hybridoma protocol (46) and the immunization and fusion, a series of ELISA screens were performed using a peptide antigen titration from 1 × 10−7 to 1 × 10−10 M to ensure that only the hybridoma clones with the highest affinity (for example, able to detected antigen at a concentration less than 1 × 10−8 M) to peptide antigens were selected. IgG mAbs were selected using a Sigma antibody isotyping kit (no. 11493027001). Four to six IgG hybridomas per peptide antigen were selected for multiple rounds of limited dilution subcloning to ensure stability and monoclonality. Each hybridoma cell line was used to prepare milliliters of ascites containing 1 to 10 mg of mouse IgGs. Mouse strains used for immunization and ascite production were BALB/c and F1 from Shanghai Super-B&K Laboratory Animal Co. Ltd. The procedures for care and use of animals were approved by the Abmart Institutional Animal Care and Use Committee.

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