APPLICATION NOTES
Read about how scientists are using Alto to simplify their interaction characterizations.
Accelerate lead identification with high-precision Digital SPR analysis
Lead identification requires screening vast molecular libraries (hybridoma, phage, B-cell, or AI-generated) and thorough characterization, including epitope binning, binding kinetics, and developability assessment. Choosing the right label-free biosensor with the ideal balance of data quality, throughput, automation, and cost remains a challenge. Alto simplifies this process with high throughput, fast assay optimization, exceptional precision, and automated ease-of-use for rapid binding interaction analysis.
Discover how Alto empowers you by providing a unique system with high-throughput, fast assay optimization ability, excellent precision, and automated ease-of-use for rapid assessment of binding interactions. From small peptides to antibodies to virus particles, Alto’s automated workflow ensures reliable and reproducible results, making it an indispensable tool for advancing pharmaceutical research.
Alto streamlines high-throughput screening with a 16-parallel sensor format and automated sample dilutions — all within a self-contained cartridge. Focus on results, not sample handling, while optimizing throughput, cost, and ease of use.
Accurately measure affinities from picomolar to micromolar levels, covering diverse ligand-antigen systems.
Get reliable KD, kon, and koff values with Alto’s digital SPR, delivering results on par with traditional SPR and BLI—without the limitations of older technologies.
Alto’s 16-parallel sensors and automated loading enable walkaway analysis of hundreds of samples. Plate handling automation enables 24×7 unattended runs.
Analyze interactions with just 2 µL of sample while Alto handles on-board 5x 3-fold dilutions—ideal for limited or costly materials.
Alto’s capture screening protocol enables rapid characterization of up to 48 ligands from complex matrices on a single cartridge. Screen 48 antibodies against one antigen in under 5 hours or two antigens in under 12—using 100X less sample. With one-click analysis, minimal maintenance, and crude sample compatibility, Alto streamlines biotherapeutic screening.
Series of sensorgrams collected for a single lane of the capture screening assay with high throughput expansion turned on. The sensorgrams show the capture (or absence of capture) of the ligand followed by washing with buffer and then binding (or absence of binding) of the antigen. Antibodies loaded in wells D1-I1 are tested against both antigens, resulting in an overlay of twelve curves for each lane of test, and 96 overall.
Alto accurately measured affinities across five orders of magnitude for eight bispecific ImmTAC® molecules, matching traditional SPR in affinity and off-rate ranking. Using a single cartridge, Alto’s SPR assay delivered results consistent with fluidics-based SPR, with differences well within expected instrument variation.
The SPR binding assay for pHLA ligand and 8 bispecific ImmTAC molecules were performed in a single cartridge with Alto. The accuracy of the kinetic and affinity results obtained by Alto was supported by running the same set of samples in a traditional fluidics-based SPR instrument. The difference between Alto kinetics values compared to traditional SPR are shown in the figures below and summarized in the table. These values are well within the expected variation when comparing between instruments, operators and methods. This accuracy in the absolute values of the kinetics and affinity also mean that the affinity and off-rate ranking is strongly in agreement with the ranking produced by the traditional SPR instrument.
| Alto | Traditional SPR | |||||
|---|---|---|---|---|---|---|
|
Analyte |
ka(M-1s-1) |
kd(s-1) |
KD(nM) |
ka(M-1s-1) |
kd(s-1) |
KD(nM) |
|
1 |
– |
– |
775 |
– |
– |
792 |
|
2 |
5.61 x 105 |
2.15 x 10-2 |
38.4 |
5.25 x 105 |
5.31 x 10-2 |
101 |
|
3 |
9.40 x 105 |
2.24 x 10-3 |
2.38 |
1.15 x 106 |
9.05 x 10-3 |
7.89 |
|
4 |
1.04 x 106 |
2.20 x 10-3 |
2.1 |
8.97 x 105 |
1.47 x 10-3 |
1.64 |
|
5 |
9.8 x 105 |
2.10 x 10-4 |
0.21 |
1.76 x 106 |
3.34 x 10-4 |
0.19 |
|
6 |
2.09 x 106 |
1.14 x 10-4 |
0.054 |
1.31 x 106 |
1.38 x 10-4 |
0.105 |
|
7 |
1.35 x 106 |
5.62 x 10-5 |
0.041 |
1.37 x 106 |
2.20 x 10-5 |
0.0161 |
|
8 |
2.39 x 106 |
2.03 x 10-5 |
0.0085 |
1.65 x 106 |
1.22 x 10-5 |
0.0074 |
Kinetic parameters measured for 8 analytes using Alto.
Alto accurately measured affinities across five orders of magnitude for eight bispecific ImmTAC® molecules, matching traditional SPR in affinity and off-rate ranking. Using a single cartridge, Alto’s SPR assay delivered results consistent with fluidics-based SPR, with differences well within expected instrument variation.
The SPR binding assay for pHLA ligand and 8 bispecific ImmTAC molecules were performed in a single cartridge with Alto. The accuracy of the kinetic and affinity results obtained by Alto was supported by running the same set of samples in a traditional fluidics-based SPR instrument. The difference between Alto kinetics values compared to traditional SPR are shown in the figures below and summarized in the table. These values are well within the expected variation when comparing between instruments, operators and methods. This accuracy in the absolute values of the kinetics and affinity also mean that the affinity and off-rate ranking is strongly in agreement with the ranking produced by the traditional SPR instrument.
| Alto | Traditional SPR | |||||
|---|---|---|---|---|---|---|
|
Analyte |
ka(M-1s-1) |
kd(s-1) |
KD(nM) |
ka(M-1s-1) |
kd(s-1) |
KD(nM) |
|
1 |
– |
– |
775 |
– |
– |
792 |
|
2 |
5.61 x 105 |
2.15 x 10-2 |
38.4 |
5.25 x 105 |
5.31 x 10-2 |
101 |
|
3 |
9.40 x 105 |
2.24 x 10-3 |
2.38 |
1.15 x 106 |
9.05 x 10-3 |
7.89 |
|
4 |
1.04 x 106 |
2.20 x 10-3 |
2.1 |
8.97 x 105 |
1.47 x 10-3 |
1.64 |
|
5 |
9.8 x 105 |
2.10 x 10-4 |
0.21 |
1.76 x 106 |
3.34 x 10-4 |
0.19 |
|
6 |
2.09 x 106 |
1.14 x 10-4 |
0.054 |
1.31 x 106 |
1.38 x 10-4 |
0.105 |
|
7 |
1.35 x 106 |
5.62 x 10-5 |
0.041 |
1.37 x 106 |
2.20 x 10-5 |
0.0161 |
|
8 |
2.39 x 106 |
2.03 x 10-5 |
0.0085 |
1.65 x 106 |
1.22 x 10-5 |
0.0074 |
Kinetic parameters measured for 8 analytes using Alto.
The unique DMF-based Alto system is revolutionizing SPR assays. The Alto is bringing unprecedented ease of use and unattended operation, while providing high-quality data for up to 48 samples per cartridge, without the need for set up manual dilutions.
Learn more about the technology and its wide application across research and drug discovery and development.
Read about how scientists are using Alto to simplify their interaction characterizations.
