Evaluation of monospecific antibodies: a comparison study with commercial analogs using immunohistochemistry on tissue microarrays.
Generation of monospecific antibodies (msAbs) (multiepitope) through affinity purification of polyclonal antisera is a plausible strategy for high-throughput production of affinity reagents toward large sets of proteins. These antibodies are generated using readily accessible gene sequence information from publicly available databases. The resulting antibodies have the potential to be used in a variety of assays, probing differentially presented and altered proteins with high sensitivity and specificity. In the present study, 48 msAbs were compared with corresponding commercial analogs. Immunohistochemical staining properties were evaluated on tissue microarrays, representing various normal human tissues from 144 different individuals. MsAbs showed similar immunostaining patterns as compared with corresponding commercial analogs in 44 out of totally 48 (92%) antibody pairs analyzed. Although only few antibody pairs showed major discrepancies, minor dissimilarities were frequently seen. Our results suggest that msAbs are reliable and valuable tools in antibody-based proteomics, enabling analysis of protein expression patterns in cells and tissues. High-throughput strategies employing such antibodies provide a consistent approach in the exploration of the human proteome.
Paavilainen
Linda
L
Department of Genetics and Pathology, The Rudbeck Laboratory, Uppsala University, Uppsala, Sweden.
Wernérus
Henrik
H
Nilsson
Peter
P
Uhlén
Mathias
M
Hober
Sophia
S
Wester
Kenneth
K
Pontén
Fredrik
F
D000906:Antibodies; D000915:Antibody Affinity; D000918:Antibody Specificity; D015153:Blotting, Western; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D007202:Indicators and Reagents / Q000592:standards; D009928:Organ Specificity / Q000276:immunology; D040081:Protein Array Analysis; D020543:Proteome / Q000276:immunology; D040901:Proteomics; D046888:Tissue Array Analysis / Q000379:methods / Q000592:standards
D000906:Antibodies; D000939:Epitopes; D007202:Indicators and Reagents; D020543:Proteome