Antibody drugs, such as monoclonal antibodies and antibody-drug conjugates, have shown significant potential in treating diseases due to their high specificity and affinity. In vivo analysis of antibody drugs with non-invasive and real-time techniques is of importance to understand dynamic behavior of drugs within living organisms, and help evaluate their pharmacokinetics and efficacies. This review summarizes the advances and in vivo analysis methods of antibody drugs, including the techniques of radiolabeled imaging, near-infrared fluorescence imaging and surface-enhanced Raman spectroscopy. The principles, applications, and challenges of each technique are discussed, which provides insights for the development of antibody drugs and in vivo analytical methods.