Construction of a large phage display antibody library by in vitro package and in vivo recombination.
Capacity and diversity are extremely important to the quality of various phage display libraries. In this work, lambda phage-based in vitro package was applied to construct a filamentous phage display antibody library so as to enlarge its capacity and introduce more sequence diversity in the final library. In vivo recombination via Cre recombinase/lox sites was also exploited to create V(H)/V(L) combination diversity based on multivalent package of lambda phage packaging extracts on phagemid DNA concatemers. The library constructed with 10 microg concatenated phagemid DNA and ten vials of lambda phage packaging extracts was calculated to contain 1.40 x 10(10) independent clones. Higher capacity can be easily achieved when more materials are consumed. This strategy is somewhat more efficient than prior methods.
Cen
Xiaodong
X
College of Life Sciences, Peking University, Beijing 100871, People's Republic of China.
Bi
Qun
Q
Zhu
Shenggeng
S
D000906:Antibodies / Q000235:genetics*; D010582:Bacteriophage lambda / Q000235:genetics*; D022801:Complementarity Determining Regions / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000821:virology*; D005822:Genetic Vectors; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D019151:Peptide Library; D011995:Recombination, Genetic
D000906:Antibodies; D022801:Complementarity Determining Regions; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D019151:Peptide Library