Unknown

Dataset Information

0

Automated screening of monoclonal antibodies for SISCAPA assays using a magnetic bead processor and liquid chromatography-selected reaction monitoring-mass spectrometry.


ABSTRACT: Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA) utilizes antibodies to enrich peptides from complex matrices for quantitation by stable isotope dilution mass spectrometry. SISCAPA improves sensitivity and limits the sample handling required for plasma-based analysis. Thus far, SISCAPA assays have been performed using polyclonal antibodies, yet monoclonal antibodies are an attractive alternative since they provide exquisite specificity, a renewable resource, and the potential for isolation of clones with very high affinities (10(-9) M or better). The selection of a good monoclonal antibody out of hundreds-to-thousands of clones presents a challenge, since the screening assay should ideally be in the format of the final SISCAPA assay, but performing the assays manually is labor- and time-intensive. In this manuscript, we demonstrate that monoclonal antibodies can be used in SISCAPA assays, and we describe an automated high-throughput SISCAPA method that makes screening of large numbers of hybridomas feasible while conserving time and resources.

SUBMITTER: Schoenherr RM 

PROVIDER: S-EPMC2839902 | BioStudies | 2010-01-01

REPOSITORIES: biostudies

Similar Datasets

2009-01-01 | S-EPMC2689780 | BioStudies
2012-01-01 | S-EPMC3253882 | BioStudies
2015-01-01 | S-EPMC4552657 | BioStudies
2012-01-01 | S-EPMC3317677 | BioStudies
1000-01-01 | S-EPMC2808264 | BioStudies
2017-01-01 | S-EPMC5540962 | BioStudies
2009-01-01 | S-EPMC2865473 | BioStudies
1986-01-01 | S-EPMC1147009 | BioStudies
2017-01-01 | S-EPMC5629267 | BioStudies
2013-01-01 | S-EPMC3563841 | BioStudies