Preparation of monoclonal antibody to human augmenter of liver regeneration:screening of hybridomas with unpurified antigen expressed by E.coli / 中国免疫学杂志

Objective:To prepare monoclonal antibody(McAb) against human augmenter of liver regeneration(hALR) by hybridoma technique with unpurified recombinant protein expressed by E.coli as screening antigen.Methods:BALB/c mice were immunized with hALR thioredoxin(hALR Thi) fusion protein twice.The splenocytes were fused with myeloma cells by polyethylene glycol and the hybridomas were selected in HAT medium.The positive hybridomas secreting anti hALR McAb were screened by means of ELISA,with the lysates of E.coli containing the recombinant plasmid pQE30 hALR induced by IPTG as screening antigen and containing plasmid pQE30 as controlling antigen.The reactivity of McAb secreted by the positive hybridoma to hALR expressed in eukaryocyte and native hALR in human serum was confirmed by means of ELISA and Western blot methods.Results:A positive hybridoma was screened successfully;the specific antigen antibody reaction was confirmed between the McAb prepared and hALR expressed in eukaryocyte and native hALR in human serum.Conclusion:Without any purification procedure,recombinant protein expressed by E.coli can be used for screening bybridomas in preparing of McAb with the expressed product of empty plasmid as control;anti hALR McAb can be used to the further study of hALR.

Study of the application of scleral flap made by tunnel knife in trabeculectomy / 重庆医科大学学报

Objective:To study the effect and complication of trabeculectomy with scleral flap made by tunnel knife in prospective randomized clinical trial.Methods:A total of 40 glaucoma eyes were divided into two groups by randomized method,Trabeculectomy was performed with fornix-based flaps,and scleral flaps made by tunnel knife(group A);or limbus-based flaps,and scleral flaps made by general method(group B)respectively.The samples were observed for 6 months after operation and the effect and complication of the two groups were compared.Results:There was no significant difference between group A and B in success rate,which were 75% and 70% respectively;and the conditiong of complication was the same.Conclusion:It can effectively simplify the procedure of fornix-based flaps trabeculectomy with scleral flaps made by tunnel knife.