Rapid antibody quantification and generation of whole proteome antibody response profiles using LIPS (luciferase immunoprecipitation systems).
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.
Burbelo
Peter D
PD
Department of Oncology, Georgetown University Medical Center, Washington, DC 20057, USA. burbp1@yahoo.com
Ching
Kathryn H
KH
Mattson
Thomas L
TL
Light
Jason S
JS
Bishop
Lisa R
LR
Kovacs
Joseph A
JA
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
D000906:Antibodies; D016655:HIV Core Protein p24; D020543:Proteome; D008156:Luciferases