Antibodies for proteomic research: comparison of traditional immunization with recombinant antibody technology.
Antibodies play a pivotal role in studying the expression and function of proteins. Proteomics studies require the generation of specific and high-affinity antibodies against large numbers of proteins. While traditional animal-based antibody generation is laborious, difficult to automate, and therefore less suited to keep up with the requirements of proteomics research, the use of recombinant in vitro antibody technology might offer a solution to this problem. However, it has not been demonstrated yet that such antibodies are at least as useful as conventional antibodies for typical proteomics applications. Here we generated novel recombinant Fab antibody fragments from the naïve HuCAL GOLD library against a number of targets derived from a mouse cDNA library. We compared these antibodies with polyclonal antisera produced against the same targets and show that these recombinant antibodies are useful reagents for typical applications like Western blotting or immunohistochemistry.
Ohara
Reiko
R
Kazusa DNA Research Institute, Kisarazu, Chiba, Japan.
Knappik
Achim
A
Shimada
Kiyo
K
Frisch
Christian
C
Ylera
Francisco
F
Koga
Hisashi
H
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D015153:Blotting, Western / Q000379:methods; D001921:Brain / Q000276:immunology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D040901:Proteomics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology*; D014611:Vaccination
D007140:Immunoglobulin Fab Fragments; D011993:Recombinant Fusion Proteins