Efficient construction of a large nonimmune phage antibody library: the production of high-affinity human single-chain antibodies to protein antigens.
A large library of phage-displayed human single-chain Fv antibodies (scFv), containing 6.7 x 10(9) members, was generated by improving the steps of library construction. Fourteen different protein antigens were used to affinity select antibodies from this library. A panel of specific antibodies was isolated with each antigen, and each panel contained an average of 8.7 different scFv. Measurements of antibody-antigen interactions revealed several affinities below 1 nM, comparable to affinities observed during the secondary murine immune response. In particular, four different scFv recognizing the ErbB2 protein had affinities ranging from 220 pM to 4 nM. Antibodies derived from the library proved to be useful reagents for immunoassays. For example, antibodies generated to the Chlamydia trachomatis elementary bodies stained Chlamydia-infected cells, but not uninfected cells. These results demonstrate that phage antibody libraries are ideally suited for the rapid production of panels of high-affinity mAbs to a wide variety of protein antigens. Such libraries should prove especially useful for generating reagents to study the function of gene products identified by genome projects.
Sheets
M D
MD
Department of Molecular Cell Biology, University of California, Berkeley, CA 94720, USA.
Amersdorfer
P
P
Finnern
R
R
Sargent
P
P
Lindquist
E
E
Schier
R
R
Hemingsen
G
G
Wong
C
C
Gerhart
J C
JC
Marks
J D
JD
Lindqvist
E
E
D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000276:immunology*; D000915:Antibody Affinity; D000941:Antigens / Q000276:immunology*; D015723:Gene Library; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology*; D051379:Mice; D011506:Proteins / Q000276:immunology*
D000906:Antibodies; D000941:Antigens; D007135:Immunoglobulin Variable Region; D011506:Proteins