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. Author manuscript; available in PMC: 2012 Feb 1.
Published in final edited form as: J Immunol Methods. 2010 Nov 13;364(1-2):50–64. doi: 10.1016/j.jim.2010.11.001

Figure 5.

Figure 5

RabMAb selection by MiSCREEN and comparison to SPR. Panel A: Hybridoma supernatants containing RabMAbs specific for thyroglobulin peptide CPTAC-39d were screened by MiSCREEN using a wash time of 10 minutes. The same supernatants were also tested for peptide binding kinetics by SPR. The vertical axis represents the accumulated MALDI peak intensity for each RabMAb while the horizontal axis represents the dissociation constant (kd) for the same antibody. Antibodies that had a high enough affinity to retain the target peptide during the wash steps (approximately 10 mins; kd of ~1.0E-03) showed a high MALDI signal by MiSCREEN. Panel B: Hybridoma supernatants containing RabMAbs specific for thryoglobulin peptide CPTAC-39c were screened by MiSCREEN using a wash time of approximately 4 minutes (kd of ~3.5E-03). Antibodies with off-rates below this threshold performed weakly in the MiSCREEN assay while high affinity RabMAbs capable of retaining peptides beyond the wash period yielded a strong MALDI signal. The vertical lines in each panel represent the half off-times (10 minutes in Panel A and 4 minutes in Panel B).