Dynamic tracking analysis of monoclonal antibodies (mAbs) biotransformation in vivo is crucial, as certain modifications could inactivate the protein and reduce drug efficacy. However, a particular challenge (i.e. immune recognition deficiencies) in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen. To address this limitation, a multi-epitope affinity technology utilizing the metal organic framework (MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region (CDR) mimotope peptide (HH24) and non-CDR mimotope aptamer (CH1S-6T) onto the surface of MOF@Au nanocomposite. Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited significantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope affinity technology. Moreover, the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HC-Asn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry (LC-QTOF-MS). Therefore, multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb. Particularly, the novel bioanalytical platform can be successfully used for the tracking analysis of trastuzumab modifications in different biological fluids. Compared to the spiked phosphate buffer (PB) model, faster modification trends were monitored in the spiked serum and patients' sera due to the catalytic effect of plasma proteins and relevant proteases. Differences in peptide modification levels of trastuzumab in patients' sera were also monitored. In summary, the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb, contributing to improved understanding and paving the way for future research and clinical applications.

Objective To study the expressions of PTTG and bFGF proteins and their relationship with microvessels density(MVD)in esophageal carcinoma.Methods Immunohistochemical SP method was used to detect the expression of PTTG and bFGF proteins in 48 esophageal carcinoma tissues and the same para-cancerous tissues.MVD was evaluated by immunohistochemieal staining with antibody CD34.Results The positive rate of PTTG and bFGF was 68.8%(33/48)and 70.8%(34/48)respectively.Rate of PTTG protein expression in esophageal carcinoma tissues was significantly higher than that in para-cancerous tissues(8.3%and 12.5%,P＜0.05).The positive rate 0f PTTG,bFGF and MVD was correlated with lymph node metastasis and TNM stage.There was no relationship with age,sex,tumor size MVD(P＜0.05).Conclusion PTTG and bFGF are over-expressed in esophageal carcinoma.Increased PTTG may play an important role in carcinogenesis and development of esophageal carcinoma by promoting the expression of bFGF protein which may induce an angiogenesis.