To make further investigation of the IgE antibodyrepertoire in Trichosanthin (TCS) allergic responses, amurine IgE phage surface display library was constructed(3.0×105 independent clones). We first constructed t...To make further investigation of the IgE antibodyrepertoire in Trichosanthin (TCS) allergic responses, amurine IgE phage surface display library was constructed(3.0×105 independent clones). We first constructed theVe cDNA library (4.6×105 independent clones) and VκcDNA library (3.0×105 independent clones). Then, theVε and Vκ gene segments were amplified from both libraries by PCR respectively, and assembled into Fab fragment by SOE PCR. The phage library containing Fabs wasthus constructed. The diversity of Vε from this library wasanalyzed and proved. Fab clones with high specificity toTCS have been screened out.展开更多
Recently we constructed a murine IgE phage surfacedisplay library and screened out two IgE (Fab) cloneswith specific binding activity to Trichosanthin (TCS). Inthis work, the Vε and Vκ genes of the two clones werese...Recently we constructed a murine IgE phage surfacedisplay library and screened out two IgE (Fab) cloneswith specific binding activity to Trichosanthin (TCS). Inthis work, the Vε and Vκ genes of the two clones weresequenced and their putative germline gene usages werestudied. On the basis of the known 3D structure of Trichosanthin and antibody, molecular modeling was carriedout to study the antigen-antibody interaction. The possible antigenic determinant sites on the surface of TCSrecognized by both the clones were analyzed, and the reaction forces between TCS and two Fab fragments werealso analyzed respectively.展开更多
文摘To make further investigation of the IgE antibodyrepertoire in Trichosanthin (TCS) allergic responses, amurine IgE phage surface display library was constructed(3.0×105 independent clones). We first constructed theVe cDNA library (4.6×105 independent clones) and VκcDNA library (3.0×105 independent clones). Then, theVε and Vκ gene segments were amplified from both libraries by PCR respectively, and assembled into Fab fragment by SOE PCR. The phage library containing Fabs wasthus constructed. The diversity of Vε from this library wasanalyzed and proved. Fab clones with high specificity toTCS have been screened out.
文摘Recently we constructed a murine IgE phage surfacedisplay library and screened out two IgE (Fab) cloneswith specific binding activity to Trichosanthin (TCS). Inthis work, the Vε and Vκ genes of the two clones weresequenced and their putative germline gene usages werestudied. On the basis of the known 3D structure of Trichosanthin and antibody, molecular modeling was carriedout to study the antigen-antibody interaction. The possible antigenic determinant sites on the surface of TCSrecognized by both the clones were analyzed, and the reaction forces between TCS and two Fab fragments werealso analyzed respectively.
