title: [Construction of personalized full-length fully human mammalian display antibody library for children with systemic lupus erythematosus].
abstract: OBJECTIVE
  To construct a personalized full-length fully human antibody mammalian display library for children with systemic lupus erythematosus (SLE).
  
  
  METHODS
  The total RNA was isolated from the PBMCs of SLE children. The heavy chain variable region and kappa light chain (VH and LCκ) of the antibody genes were amplified by RT-PCR and inserted into the pDGB-HC-TM vector separately to construct the heavy chain and light chain libraries. The library DNAs were transfected into 293T cells and the expression of full-length fully human antibody on the surface of 293T cells was analyzed by flow cytometry.
  
  
  RESULTS
  Using 0.8 µg total RNA as the template, the VH and LCκ were amplified and the full-length fully human antibody mammalian display library was constructed. The VH and LCκ gene libraries had a size of 9.4×10(4) and 8.4×10(4), respectively. Sequence analysis of 10 clones randomly selected from the VH and LCκ gene libraries each showed that 8 heavy chain clones and 7 light chain clones contained correct open reading frames, and flow cytometry demonstrated that all the 15 clones express full-length antibodies on 293T cell surfaces. 293T cells co-transfected with the VH and LCκ gene libraries expressed the full-length antibodies on the cell surface.
  
  
  CONCLUSIONS
  The personalized full-length fully human antibody library for SLE children constructed allows display of the full-length antibodies on mammalian cell surfaces, thus providing a valuable platform for analyzing the autoantibodies, their etiological role, and their clinical implications in SLE.
authors.affiliation: Department of Pediatrics, Second Affiliated Hospital of Guangzhou Medical College, Guangzhou , China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Zhou
authors.forename: Meihua
authors.initials: M
authors.lastname: Zhu
authors.forename: Zhongkun
authors.initials: Z
authors.lastname: Liang
authors.forename: Zhenrui
authors.initials: Z
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: He
authors.forename: Changzheng
authors.initials: C
authors.lastname: Li
authors.forename: Wanlong
authors.initials: W
authors.lastname: Tan
authors.forename: Shibo
authors.initials: S
authors.lastname: Jiang
authors.forename: Shuwen
authors.initials: S
authors.lastname: Liu
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhou
authors.forename: Chen
authors.initials: C
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D002648:Child; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D008180:Lupus Erythematosus, Systemic / Q000235:genetics* / Q000276:immunology*; D008565:Membrane Proteins / Q000235:genetics
chemical_list: D007143:Immunoglobulin Heavy Chains; D007145:Immunoglobulin kappa-Chains; D008565:Membrane Proteins