Construction and identification of non-immunized human phage display library / 中国免疫学杂志
Chinese Journal of Immunology
;
(12)1985.
Artigo
em Chinês
| WPRIM
| ID: wpr-548769
ABSTRACT
Objective:
To develop non-immunized human phage display library.Methods:
The total RNA of lymphocyte cells from peripheral blood of healthy voluntee was isolated and cDNA was synthesized,and the genes of heavy variable chain (VH) and light variable chain (V? and V?) were amplified by direct PCR and half-nested PCR.By overlapping extension PCR,the genes of VH and VL (V? and V?) were linked.The linked genes of single chain Fv fragment (scFv) were ligated with the vector pCANTAB-5E and then cloned into TG1 for the scFv library construction.Results:
By direct PCR and half-nested PCR,42 VH fragments,16 V? and 18 V? fragments were obtained.The size of linked scFv library genes was 750 bp and the volume of constructed scFv library was 1.35?108.The results of BstN Ⅰ analysis of scFv genes from the phage library showed that fingerprint map of the selected scFvs was different.Conclusion:
The developed phage library is diversity and can be used for selecting humanized scFv.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Tipo de estudo:
Estudo diagnóstico
/
Estudo prognóstico
Idioma:
Chinês
Revista:
Chinese Journal of Immunology
Ano de publicação:
1985
Tipo de documento:
Artigo
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