Development of recombinant anti-human CD antibodies for diabetes and cancer monitoring using flow cytometry

TBNK cells, namely T, B, Natural Killer (NK) cells, are three major types of lymphocytes in the immune system. Comprehensive analysis of cluster of differentiation (CD) antigens on TBNK cell surface, such as CD3, CD4, CD8, CD16, CD19, CD56, and CD45, contributes to precise immune cell profiling and disease progression tracking. In this study, we prepared recombinant antibodies specifically target CDs markers using the ExpiCHO-Double Plasmid system, which ensured high-yield expression and reproducibility. The antibodies were efficiently conjugated with various fluorescent dyes, including FITC, PE-Cy7, APCCy7, APC, PE, and PerCP-Cy5.5, to achieve multiplexed flow cytometry analysis of TBNK cell populations. Testing of clinical samples demonstrated that the developed antibody conjugates could effectively distinguish the difference of TBNK cell populations between healthy individuals and patients, analysis of 291 adults showed type 2 diabetes had elevated CD4⁺/CD8⁺ ratios (1.9x) versus reduced ratios in cancer (0.75x), with principal component analysis (PCA) distinguishing groups (PC1 = 47.9 %) indicating their potential for disease monitoring and therapeutic evaluation. Our results underscore the utility of novel immunodiagnostic tools for early detection and personalized management of diabetes and cancer.