Determination of functional affinity of rhEndoglin conjugated eptide / 第三军医大学学报

Objective To assess functional affinity of rhEndoglin conjugated eptide in order to identify the affinity.Methods We measured the functional affinity of rhEndoglin conjugated eptide with non-competitive ELISA method.After coating the peptide by BSA binding with glutaral couple,affirming the best concentration,best time of peptide to coat the plate and the coating coefficient,and the proper binding time of peptide to Endoglin to reach an equilibrium,we plotted the standard curve of the binding reaction of peptide and Endoglin.Results The affinity constant of peptide and anti-hEndoglin IgG is respectively(2.956?0.749)?106mol/L and(7.403?10.76) ?108mol/L.Conclusion The peptide we got from the peptide liberary can strongly bind to Endoglin,providing a theoretical basis for its clinical use.

Screening of conjugated peptides of Endoglin from phage display peptide library / 第三军医大学学报

Objective To screen rhEndoglin-binding peptides from phage displayed 12-peptide library. Methods The rhEndoglin was used as target protein for biopanning of phage-displayed 12-peptide library. After three rounds of screening, 16 phage clones were randomly selected and identified by sandwich ELISA. The positive phage clones were sequenced, and the fuse peptides were deduced by the DNA sequence. Further we identified the affinity and speciality by competitive inhibition test. Results Six of 16 phage clones were identified as positive clones by competent ELISA which could bind to rhEndoglin. Five sequences were obtained, and the amino acid sequence in two of these five was AHKHVHHVPVRL. Conclusion The rhEndoglin-binding peptides can be obtained by screening phage random peptide library. It could play an important role in early diagnosis of ovarian cancer.

Screening and function analysis of targeted short binding peptides of Endoglin / 重庆医科大学学报

Objective:To do screening in phage displayed 12-peptide library to seek short peptides capable of binding Endoglin detect their affinity constants.Methods:After three rounds of screening,16 phage clones were randomly selected and their affinity was identified by sandwich ELISA.The DNA sequence of positive phage clones was determined,and their speciality was identified by competitive inhibition test.We finally calculated out the affinity constants of those positive phage binding peptides by non-competitive ELISA method.Result:After three rounds of screening,the enriched phage clones were identified. 6 of 16 phage clones were identified positive by competitive ELISA,which had comparatively strong binding activity to rhEndoglin.Five sequences were obtained,and the predominant sequence was AHKHVHHVPVRL.Competitive inhibition test showed that positive phage clones had good affinity to Endoglin,with the affinity constant as(1.431?0.293)?107 mol/L.Conclusion:The rhEndoglin-binding peptides with high affinity to Endoglin can be obtained through the screening of phage random peptide library, which is beneficial to the further studies on the function of Endoglin in the early diagnosis of ovarian cancer.