Construction and diversity analysis of a murine IgE phage surface display library.
To make further investigation of the IgE antibody repertoire in Trichosanthin (TCS) allergic responses, a murine IgE phage surface display library was constructed (3.0 x 10(5) independent clones). We first constructed the V epsilon cDNA library (4.6 x 10(5) independent clones) and V kappa cDNA library (3.0 x 10(5) independent clones). Then, the V epsilon and V kappa gene segments were amplified from both libraries by PCR respectively, and assembled into Fab fragment by SOE PCR. The phage library containing Fabs was thus constructed. The diversity of V epsilon from this library was analyzed and proved. Fab clones with high specificity to TCS have been screened out.
Li
Z D
ZD
Shanghai Institute of Cell Biology, Chinese Academy of Sciences, China.
Yeh
M
M
D000818:Animals; D000916:Antibody Diversity / Q000235:genetics*; D017104:Bacteriophage M13 / Q000235:genetics* / Q000276:immunology*; D018076:DNA, Complementary / Q000032:analysis; D015723:Gene Library; D005822:Genetic Vectors; D007073:Immunoglobulin E / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007139:Immunoglobulin epsilon-Chains / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D015978:Trichosanthin / Q000276:immunology
D018076:DNA, Complementary; D007135:Immunoglobulin Variable Region; D007139:Immunoglobulin epsilon-Chains; D007145:Immunoglobulin kappa-Chains; D007073:Immunoglobulin E; D015978:Trichosanthin