ABSTRACT
OBJECTIVE@#To prepare monoclonal antibody (MAb) against carboxy-terminus of E-phA4 and to analyze the immunological characteristics and significance of the antibody.@*METHODS@#Mice were immunized with a chemically synthesized peptide that had been conjugated to KLH (keyhole limpet hemocyamin). A panel of antibodies specifically against EphA4 were obtained by hybridoma technique. The immunological properties and significance of the MAb were analyzed by immunological and immunochemistry techniques.@*RESULTS@#A hybridoma cell line secreting anti-EphA4 MAb was established. The MAb reacted specifically with human, chicken and mouse EphA4, but did not cross-react with EphA5 and EphA7. The antibody could be used for immunochemical techniques such as ELISA, immunoprecipitation, Western blot and immunohistochemistry.@*CONCLUSION@#The anti-EphA4 MAb generated by immunizing a synthetic peptide possesses excellent immunological properties. MAb can be applied for immunological and immunohistochemical purpose and will become an important tool in the study of EphA4 and its ligand.
Subject(s)
Animals , Mice , Antibodies, Monoclonal , Hybridomas , Mice, Inbred BALB C , Peptide Fragments , Allergy and Immunology , Receptor, EphA4 , Allergy and ImmunologyABSTRACT
Objective To clarify the correlation of alpha fetoprotein （AFP） half-life （T1／2） with relapse or／and metastasis of primary hepatocellular carcinoma （PHC） after treatment. Methods Determination of AFP content in serum with two steps double McAb sandwich ELISA.Calculation AFP T1／2 of the sufferers treated with intervention and radiofrequency ablation （RFA） according to T1／2=0.3 dT／log（AFP1／AFP2）. Results When AFP T1／2＞11.5 days， the sensitivity， specificity and accuracy of predicting the relapse or metastasis of PHC after treatment were 87.5％， 89.3％， 88.5％， respectively. Intervention group was 82.35％， 83.3％， 82.75％， respectively and RFA group was 93.3％， 93.75％， 93.5％， respectively. During 200 days，there was very significant difference of AFP T1／2 （P＜0.001） between non-relapsed／ non-metastasized and relapsed， metastasized and died. Conclusions AFP T1／2 can be used as a predictive index for relapse， metastasis of PHC after treatment. It is not only a reliable parameter for the later stage treatment， but also a useful index for the evaluation of effect.