Protein microarray-based screening of antibody specificity.
The increased use of antibodies as therapeutics, as well as the growing demand for large numbers of antibodies for high-throughput protein analyses, has been accompanied by a need for more specific antibodies. An array containing every protein for the relevant organism represents the ideal format for an assay to test antibody specificity since it allows the simultaneous screening of thousands of proteins in relatively normalized quantities. Indeed, the use of a yeast proleome array to profile the specificity of several antibodies directed against yeast proteins has recently been described. In this chapter, we present a detailed description of the methods used to probe protein arrays with antibodies as well as the technical issues to consider when carrying out such experiments.
Bangham
Rhonda
R
Research and Development, Protometrix Inc., Branford, CT, USA.
Michaud
Gregory A
GA
Schweitzer
Barry
B
Predki
Paul F
PF
D000906:Antibodies / Q000378:metabolism; D000918:Antibody Specificity; D005656:Fungal Proteins / Q000032:analysis; D006801:Humans; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D020543:Proteome / Q000032:analysis; D012984:Software
D000906:Antibodies; D005656:Fungal Proteins; D020543:Proteome