Protein microarrays for parasite antigen discovery.
The host serological profile to a parasitic infection, such as schistosomiasis, can be used to define potential vaccine and diagnostic targets. Determining the host antibody response using traditional approaches is hindered by the large number of putative antigens in any parasite proteome. Parasite protein microarrays offer the potential for a high-throughput host antibody screen to simplify this task. In order to construct the array, parasite proteins are selected from available genomic sequence and protein databases using bioinformatic tools. Selected open reading frames are PCR amplified, incorporated into a vector for cell-free protein expression, and printed robotically onto glass slides. The protein microarrays can be probed with antisera from infected/immune animals or humans and the antibody reactivity measured with fluorophore labeled antibodies on a confocal laser microarray scanner to identify potential targets for diagnosis or therapeutic or prophylactic intervention.
Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Herston, QLD, Australia.
Patrick
P
Driguez
Denise L
DL
Doolan
Douglas M
DM
Molina
Alex
A
Loukas
Angela
A
Trieu
Phil L
PL
Felgner
Donald P
DP
McManus
D000818:Animals; D000917:Antibody Formation / Q000276:immunology; D000953:Antigens, Protozoan / Q000276:immunology*; D030562:Databases, Protein; D005822:Genetic Vectors; D016366:Open Reading Frames; D010271:Parasites / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D040081:Protein Array Analysis / Q000379:methods*; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology; D016053:RNA, Protozoan / Q000302:isolation & purification
D000953:Antigens, Protozoan; D015800:Protozoan Proteins; D016053:RNA, Protozoan