Label-Free Assays for High-Throughput Monoclonal Antibody ...
Label-Free Assays for High-Throughput Monoclonal Antibody ...
Label-Free Assays for High-Throughput Monoclonal Antibody ...
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Typical Cross-Competition Assay Formats - (ELISA, Biacore, Octet, etc.)<br />
Classical<br />
Sandwich Assay<br />
Premix<br />
Assay<br />
In Tandem<br />
Assay<br />
• Need good quality homogeneous<br />
reagents<br />
• Antigen dissociation from mAb-1<br />
• Use anti-hFc or anti-mFc or anti-Fab<br />
surface to capture mAb-1. Need to<br />
quench unsaturated capture<br />
surface.<br />
• Use new sensor surface or optimize<br />
regeneration condition<br />
• Works <strong>for</strong> heterogeneous & dimeric<br />
antigens<br />
• Need lot of reagents<br />
mAb-1 (Saturating mAb)<br />
• Use anti-hFc or anti-mFc or anti-Fab<br />
surface to capture mAb-1. Need to<br />
quench unbound capture surface.<br />
mAb-2 (Competing mAb)<br />
• Use new sensor surface or optimize<br />
regeneration condition<br />
Target / Antigen<br />
• Works <strong>for</strong> heterogeneous reagents<br />
• Tagged antigen or directly couple to<br />
the surface<br />
• Influenced by the size of antigen<br />
• Capturing the antigen might block the<br />
epitope <strong>for</strong> mAb binding<br />
• Use new sensor surface or optimize<br />
regeneration condition<br />
Abdiche, Y. N. et al. (2012) J. Immunol.<br />
Methods 382(1-2), pp. 101-116<br />
Confidential<br />
VISHAL KAMAT
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