Ultrafine-tricalcium phosphate(β-TCP)powders with good crystalline structure were produced by a new process through bone tissue engineering approach rorous β-TCPcermic was combined with recombined human bone morphog...Ultrafine-tricalcium phosphate(β-TCP)powders with good crystalline structure were produced by a new process through bone tissue engineering approach rorous β-TCPcermic was combined with recombined human bone morphogenetic proteins-2(rhBMP-2)to develop a novel composite material ,osteogenesis capacity of the composite was investigated intramuscularly in rat with histological analyses and SEM examination pureβ-TCP porous carmic wsa investigated as the control results show that the compostie materials possess good bilcompatibility biodegradation and strong osteogenesis capacity through inductive process after implantation material degradation began from 2 weeks post-implantation accompanying with the changing o pore structure with the enwrapping and separation fo materials by hyperplatic mesenchymal cells and fibroblast and with the phagocytose reaction of multinucleated giant cells early in 72h immature cartilage could be found within novel composite mature lamellar bone was induced to generate after 3 weeks with strong osteoinduction capacity and controlable bildegradation the novel rhBMP-2\β-TCP porous ceramic is expected to be a promising bone grafting substitute for bone tissue engineering展开更多
To construct ScFv and Fab from murine anti gastric cancer monoclonal antibody(mAb)3H11. Methods.At first,3H11 ScFv and Fab were constructed with V genes PCR amplified by degenerate primers for FR1.The bacterial expres...To construct ScFv and Fab from murine anti gastric cancer monoclonal antibody(mAb)3H11. Methods.At first,3H11 ScFv and Fab were constructed with V genes PCR amplified by degenerate primers for FR1.The bacterial expressed 3H11 Ab fragments showed no antigen binding activity.Then,phage antibody library and random mutated library were constructed from 3H11 hybridoma cells and panning selection was performed.Again the identification of positive clone was failed.Finally the N terminal sequences of V regions were resumed to 3H11 original sequences by site directed mutagenesis via PCR. Results.Binding activity to gastric cancer cells was detected only from N terminal sequence corrected 3H11 ScFv and Fab,though the expression of the Ab fragments was not affected.Correction of either VL or VH N terminal sequences could partially resume the antigen binding activity. Conclusion.Sequence changes of V region N terminal introduced by PCR may seriously affect antigen binding without affecting the expression of antibody.展开更多
基金This study was financially supported by 863 Hi-Tech Research and Development Program of China(2002AA326080)The Fund for Youth Teacher of Education Ministry of China(2002123).
文摘Ultrafine-tricalcium phosphate(β-TCP)powders with good crystalline structure were produced by a new process through bone tissue engineering approach rorous β-TCPcermic was combined with recombined human bone morphogenetic proteins-2(rhBMP-2)to develop a novel composite material ,osteogenesis capacity of the composite was investigated intramuscularly in rat with histological analyses and SEM examination pureβ-TCP porous carmic wsa investigated as the control results show that the compostie materials possess good bilcompatibility biodegradation and strong osteogenesis capacity through inductive process after implantation material degradation began from 2 weeks post-implantation accompanying with the changing o pore structure with the enwrapping and separation fo materials by hyperplatic mesenchymal cells and fibroblast and with the phagocytose reaction of multinucleated giant cells early in 72h immature cartilage could be found within novel composite mature lamellar bone was induced to generate after 3 weeks with strong osteoinduction capacity and controlable bildegradation the novel rhBMP-2\β-TCP porous ceramic is expected to be a promising bone grafting substitute for bone tissue engineering
文摘To construct ScFv and Fab from murine anti gastric cancer monoclonal antibody(mAb)3H11. Methods.At first,3H11 ScFv and Fab were constructed with V genes PCR amplified by degenerate primers for FR1.The bacterial expressed 3H11 Ab fragments showed no antigen binding activity.Then,phage antibody library and random mutated library were constructed from 3H11 hybridoma cells and panning selection was performed.Again the identification of positive clone was failed.Finally the N terminal sequences of V regions were resumed to 3H11 original sequences by site directed mutagenesis via PCR. Results.Binding activity to gastric cancer cells was detected only from N terminal sequence corrected 3H11 ScFv and Fab,though the expression of the Ab fragments was not affected.Correction of either VL or VH N terminal sequences could partially resume the antigen binding activity. Conclusion.Sequence changes of V region N terminal introduced by PCR may seriously affect antigen binding without affecting the expression of antibody.
