Microarrays based on affinity-tagged SCFV antibodies: Sensitive detection of analyte in complex proteomes

• Antibody microarray is a novel technology that will allow us to efficiently perform global proteome analysis. Performing rapid and multiplexed analysis on complex proteomes, such as human serum, targeting also low-abundant analytes will, however, place high demands upon the microarray design with respect to logistics, specificity and sensitivity. In this study, we have shown that non-purified affinity-tagged scFv antibody fragments could be directly applied as probes thereby eliminating the need for any time-consuming pre-purification steps. Further, highly functional arrays were obtained providing an assay sensitivity in the pM to fM range. In fact, 300 zeptomole analyte (approx. 5,000 molecules) may be sufficient for detection. Moreover, highly complex proteomes could be analysed without impairing the specificity and sensitivity of the setups. This is a pre-requisite for the design of high-density antibody arrays applied in high-throughput proteomics.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Immunologi inom det medicinska området (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Immunology in the medical area (hsv//eng)

Nyckelord

scFv

antibody microarrays

antibody chips

sensitivity

affinity-tag

Publikations- och innehållstyp

kon (ämneskategori)

ref (ämneskategori)