Peptides and Anti-peptide Antibodies for Small and Medium Scale Peptide and Anti-peptide Affinity Microarrays: Antigenic Peptide Selection, Immobilization, and Processing.
This chapter describes the principles of selection of antigenic peptides for the development of anti-peptide antibodies for use in microarray-based multiplex affinity assays and also with mass-spectrometry detection. The methods described here are mostly applicable to small to medium scale arrays. Although the same principles of peptide selection would be suitable for larger scale arrays (with 100+ features) the actual informatics software and printing methods may well be different. Because of the sheer number of proteins/peptides to be processed and analyzed dedicated software capable of processing all the proteins and an enterprise level array robotics may be necessary for larger scale efforts. This report aims to provide practical advice to those who develop or use arrays with up to ~100 different peptide or protein features.
School of Biological Sciences, Centre for Biomedical Sciences, Royal Holloway University of London, Egham Hill, Egham, Surrey, TW20 0EX, UK.
Fan
F
Zhang
School of Biological Sciences, Centre for Biomedical Sciences, Royal Holloway University of London, Egham Hill, Egham, Surrey, TW20 0EX, UK.
Andrea
A
Briones
School of Biological Sciences, Centre for Biomedical Sciences, Royal Holloway University of London, Egham Hill, Egham, Surrey, TW20 0EX, UK. mikhail.soloviev@rhul.ac.uk.
Mikhail
M
Soloviev
D000818:Animals; D055910:Antibodies, Immobilized / Q000276:immunology*; D000941:Antigens / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D002417:Cattle; D010455:Peptides / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*; D059748:Proteolysis; D013050:Spectrometry, Fluorescence
D055910:Antibodies, Immobilized; D000941:Antigens; D010455:Peptides
Affinity assay; Antibody array; Antigenicity; Immunization; Peptide array; Peptidomics; Polyclonal antibodies; Proteomics