Antibody profiling by Luciferase Immunoprecipitation Systems (LIPS).
Technologies for comprehensively understanding and quantifying antibody profiles to autoantigens and infectious agents may yield new insights into disease mechanisms and may elucidate new markers to substratify disease with different clinical features and better understand pathogenesis. We have developed a highly quantitative method called Luciferase Immunoprecipitation Systems (LIPS) for profiling patient sera antibody responses to autoantigens and pathogen antigens associated with infection. Unlike ELISAs, the highly sensitive LIPS is easily implemented to survey humoral serological response profiles to different antigens in a universal format and produces dynamic antibody titer ranges up to 6 log(10) for some antigens. In these studies, quantitative profiling by LIPS of patient humoral responses against panels of antigens or even the entire proteome of some pathogens (i.e. HIV), is typically more informative than testing a single antigen by ELISA. In addition, LIPS also eliminates time and effort needed to produce highly purified antigens as well as the labor-intensive assay optimization steps needed for standard ELISAs. Here we provide a detailed protocol describing the technical aspects of performing LIPS assays for readily profiling antibody responses to single or multiple antigens.
Burbelo
Peter D
PD
Neurobiology and Pain Therapeutics Section, National Institute of Dental and Craniofacial Research, National Institutes of Health. burbelop@nidcr.nih.gov
Ching
Kathryn H
KH
Klimavicz
Caitlin M
CM
Iadarola
Michael J
MJ
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology; D019556:COS Cells; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D048790:Renilla / Q000201:enzymology / Q000235:genetics
D000906:Antibodies; D001324:Autoantigens; D049410:Luciferases, Renilla