title: Rapid antibody quantification and generation of whole proteome antibody response profiles using LIPS (luciferase immunoprecipitation systems).
abstract: The application of LIPS to the rapid quantification of antibody responses to infectious agents is described. Chimeric genes encoding pathogen antigens fused to Renilla luciferase are expressed in mammalian cells; crude extracts are prepared and, without purification, employed in immunoprecipitation assays to quantify pathogen-specific antibodies. In cross-sectional and longitudinal studies, antibody levels to the MSG-14 antigen of Pneumocystis jirovecii measured by this assay correlated well with levels previously obtained with an optimized ELISA. We also correctly predicted Hepatitis B (HBV), Hepatitis C (HCV), and HIV infection status in all but 2 of 99 assays analyzing 33 patient sera. We then used 15 HIV-encoded proteins comprising the whole HIV proteome to generate antibody response profiles for these 33 sera. Each HIV antigen was recognized by antibodies in serum from at least one HIV-infected individual. Data generated with these simple, quantitative antibody-detection assays have both clinical and research applications.
authors.affiliation: Department of Oncology, Georgetown University Medical Center, Washington, DC 20057, USA. burbp1@yahoo.com
authors.forename: Peter D
authors.initials: PD
authors.lastname: Burbelo
authors.forename: Kathryn H
authors.initials: KH
authors.lastname: Ching
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Mattson
authors.forename: Jason S
authors.initials: JS
authors.lastname: Light
authors.forename: Lisa R
authors.initials: LR
authors.lastname: Bishop
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Kovacs
mesh_terms: D000906:Antibodies / Q000097:blood* / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006678:HIV / Q000276:immunology; D016655:HIV Core Protein p24 / Q000097:blood / Q000276:immunology; D016174:Hepacivirus / Q000276:immunology; D006515:Hepatitis B virus / Q000276:immunology; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D045363:Pneumocystis carinii / Q000276:immunology; D020543:Proteome / Q000276:immunology* / Q000378:metabolism*; D013997:Time Factors
chemical_list: D000906:Antibodies; D016655:HIV Core Protein p24; D020543:Proteome; D008156:Luciferases