LIPS arrays for simultaneous detection of antibodies against partial and whole proteomes of HCV, HIV and EBV

被引:26
作者
Burbelo, Peter D. [1 ]
Bren, Kathleen E. [1 ]
Ching, Kathryn H. [1 ]
Gogineni, Emile S. [2 ]
Kottilil, Shyam [3 ]
Cohen, Jeffrey I. [4 ]
Kovacs, Joseph A. [2 ]
Iadarola, Michael J. [1 ]
机构
[1] Natl Inst Dent & Craniofacial Res, Neurobiol & Pain Therapeut Sect, Lab Sensory Biol, NIH, Bethesda, MD 20892 USA
[2] NIH, Dept Crit Care Med, Ctr Clin, Bethesda, MD 20892 USA
[3] NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA
[4] NIAID, Med Virol Sect, Infect Dis Lab, NIH, Bethesda, MD 20892 USA
关键词
HUMORAL IMMUNE-RESPONSE; PROTEIN MICROARRAYS; INFECTIOUS AGENTS; PROFILES; DISEASE; VACCINE;
D O I
10.1039/c0mb00342e
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
For many infectious agents, the detection of antibodies is critical for diagnosing, monitoring and understanding vaccine responses. To facilitate the highly quantitative and simultaneous analysis of antibodies against multiple proteins from infectious agents, we have developed Luciferase Immunoprecipitation Systems (LIPS) arrays. By configuring microtiter plates with multiple antigens and testing control and infected serum samples at one time in solution, LIPS arrays provided highly reproducible antibody titers to panels of antigens with a wide dynamic range of detection. While all serum samples showed similar positive and negative immunoreactivity with internal control antigens derived from Influenza and Renilla luciferase-alone protein, respectively, antibody titers to many HCV and HIV antigens were generally 10 to over 400-fold higher in the infected versus uninfected samples. Additional screening of 18 proteins from the EBV proteome with serum samples from healthy EBV-infected individuals showed statistically significant antibody titers to 50% of the proteins tested. Antibody titers for the different EBV antigens in the healthy EBV-infected individuals were markedly heterogeneous highlighting the complexity of host humoral responses. These results suggest that LIPS arrays offer a highly discriminating platform for simultaneously profiling a wide spectrum of antibodies associated with many infectious agents.
引用
收藏
页码:1453 / 1462
页数:10
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