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 theref...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2006-05, Vol.6 (9), p.2638-2646
Main Authors: Ohara, Reiko, Knappik, Achim, Shimada, Kiyo, Frisch, Christian, Ylera, Francisco, Koga, Hisashi
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Animals
Antibody
Antibody Formation
Antibody Specificity
Applied microbiology
Biological and medical sciences
Blotting, Western - methods
Brain - immunology
Fundamental and applied biological sciences. Psychology
Humans
Immunoglobulin Fab Fragments - biosynthesis
Immunoglobulin Fab Fragments - genetics
Immunoglobulin Fab Fragments - immunology
Immunohistochemistry - methods
Kinesin family member 5C
Male
Mice
Mice, Inbred ICR
Microbiology
Miscellaneous
mKIAA
MYH9
Phage display
Proteins
Proteomics
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - immunology
Vaccination
Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects)
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Summary: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.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200500579