Isolation and identification of a human single chain Fv antibody against amyloid-beta 1-42 soluble oligomers from a human phage display library / 生物工程学报

To get specific scFv (Single-chain fragment variable) antibody against soluble Abeta1-42(Amyloid-beta) oligomers, we constructed a human single-chain Fv (scFv) antibody library by phage display technology. Using RT-PCR, we amplified the variable heavy (VH) and variable light (VL) genes from peripheral blood lymphocytes (PBL). Then we obtained the scFv fragments through SOE-PCR, and the scFv fragments were cloned into the vector pCANTAB5E and electroporated into competent Escherichia coli TG1 cells. Consequently, a scFv phage display library containing 2.5 x 10(9) clones was constructed. The recombinant phagemids were rescued by reinfection of helper phage M13K07. Recombinant phages specific for Abeta1-42 oligomers were enriched after four rounds of biopanning and the antigen-positive clones were selected from the enriched clones by phage ELISA. Positive clone B19 was used to infect E. coli HB2151 to express soluble scFv antibody. SDS-PAGE and Western blotting analysis showed that the soluble scFv B19 antibody was expressed successfully and could bind specifically to Abeta1-42 trimer and protofiber. The specific scFv against Abeta1-42 oligomers can be used in the therapeutic research on Alzheimer's disease.

Degrade the Mortality Rate of Acute Gravis Type Craniocerebral Injury by Hospital Onself Quality Control / 医学研究杂志

Objective To investigate the relationship between the time factors and the remedy level for acute gravis type craniocerebral injury patients.Methods Among 367 operation cases,the preoperative time monitoring were practiced.Results Among 367 operation cases,145 cases were recovery,60 cases were medium disability,13 cases were severe handicap,10 cases were plant man,131 cases were death,8 cases self-acting discharge.Conclusions The preoperative time monitoring for acute gravis type craniocerebral injury patients and formed normalizationed standardized management institution to decurtate the preoperativ time is effective.it can elevate operation therapeutic effect,elevate subsistence quality and degrade the mortality rate and disability rate.