Table 1. Comparison of SPRi and BLI technologies with respect to throughput and sample consumption in performing a comprehensive classical sandwich epitope binning assay on 96.
Parameter | SPRi | BLI* |
Time to couple 96 ligands | 1 h (two consecutive prints of a 48-channel CFM followed by blocking of the 96-ligand array in the SPRi) | 40 min (one cycle) |
Vol. per ligand (96 total) | 100 μl | 100 μl** |
Vol. per analyte (96 total) | 120 μl | 10 ml (96× 100-μl aliquots per mAb analyte within a 384-well plate) |
Plates required to accommodate 96 mAb analytes | One 96-well plate | 24× 384-well plates |
Time to prepare 96 analytes | 30 min (dispense 100 μl per mAb into a 96-well plate) | Several hours to dispense 96 mAbs into 24 plates, each plate accommodating four mAbs, and each mAb dispensed into 96× 100 μl-aliquots |
Unattended throughput | 9216 interactions (96 analytes × 96 ligands) | 384 interactions (four analytes × 96 ligands) if one plate is used for analytes and the second plate is used for reagents. |
Unattended runtime | 30 h using standard injection times | 1 h using four 15-min cycles |
No. experiments needed | One | 24 |
Total runtime | 30 h | Several days involving 24 manual plate switches |
Reagents (antigen, buffer, and regeneration) | 10 ml each accommodated in 11-ml vials | 10 ml each, but must be distributed as 96× 100-μl aliquots into a 384-well plate |
* Robotic integration with the BLI platform results in a higher unattended throughput, longer unattended runtime, and significantly shorter total runtime by automating multi-plate analyses. Less time is needed for sample preparation if a robot is used for the microfluidic dispensing.
**Sample volumes can be reduced by 50% if tilted-bottom 384-well microplates that hold a minimum of 40 μl/well are used instead of the standard flat-bottom 384-well microplates.