A sensitive and high-throughput assay to detect low-abundance proteins in serum.
The ability to detect antigens immunologically is limited by the affinity of the antibodies and the amount of antigens. We have now succeeded in creating a modular, facile amplification system, termed fluorescent amplification catalyzed by T7 polymerase technique (FACTT). Such a system can detect protein targets specifically at subfemtomolar levels ( approximately 0.08 fM). We describe here the detection of Her2 (also known as Neu) from rodent and human sera. FACTT is adaptable to high-throughput screening and automation and provides a practical method to enhance current ELISAs in medical practice.
Pathology and Laboratory Medicine, School of Medicine, University of Pennsylvania, 3620 Hamilton Walk, Philadelphia, Pennsylvania 19104, USA.
Hongtao
H
Zhang
Xin
X
Cheng
Mark
M
Richter
Mark I
MI
Greene
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000097:blood; D061067:Antibodies, Monoclonal, Humanized; D001681:Biological Assay; D019921:Biotinylation; D001943:Breast Neoplasms / Q000097:blood* / Q000235:genetics; D002460:Cell Line; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006801:Humans; D007279:Injections, Subcutaneous; D051379:Mice; D008819:Mice, Nude; D008954:Models, Biological; D009363:Neoplasm Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D021141:Nucleic Acid Amplification Techniques; D012313:RNA / Q000032:analysis; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000097:blood* / Q000235:genetics; D011994:Recombinant Proteins / Q000097:blood; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000378:metabolism; D000068878:Trastuzumab; D047368:Tumor Burden; D014764:Viral Proteins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D012321:DNA-Directed RNA Polymerases; D005456:Fluorescent Dyes; D009363:Neoplasm Proteins; D012313:RNA; D018719:Erb-b2 Receptor Tyrosine Kinases; D011994:Recombinant Proteins; D019809:Streptavidin; D000068878:Trastuzumab; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase