Site-specific labeling of antibodies with quantum dots could promote to retain the antigen binding capacity of antibodies

A major concern with antibody labeling is the decreased antigen affinity binding capacity of antibodies, owing mainly to the randomly oriented binding of the marker. Herein, a universal approach for site-specific photocrosslinking of quantum dots (QDs) to the Fc-terminal of antibodies was investigated utilizing antibody Fc-terminal affinity proteins. Results showed that the QDs only bound to the heavy chain of the antibody. Further comparative tests confirmed that the site-specific directed labeling approach maximizes the retention of the antigen-binding capacity of the natural antibody. Compared with the commonly employed random orientation labeling approach, the directional labeling approach allows the labeled antibody showed 6 times greater binding affinity to antigen. QDs-labeled monoclonal antibodies were applied to fluorescent immunochromatographic test strips for the detection of shrimp tropomyosin (TM). The established procedure has a detection limit of 0.054 μg/mL. Thus, the site-specific labeling approach significantly improves the antigen binding capacity of the labeled antibody.